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2023-01-01 00:00:00
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10.15252/msb.20209833
|
Single-cell transcriptomics reveals immune response of intestinal cell types to viral infection
|
2021
|
Figure 1
|
<p><strong>Figure 1. Interferons protect human intestinal organoids from HAstV1 infection.</strong></p><p><strong>A</strong>. Cryo-sections of human organoids were analyzed for the presence of enterocytes (E-cad), Goblet cells (Muc-2) and tight junctions (ZO-1) by indirect immunofluorescence. Nuclei are stained with DAPI. Scale bar 25 μm.</p><p><strong>B</strong>. Human intestinal organoids were incubated with media (mock) or infected with HAstV1. 16hpi organoids were frozen, cryo-sectioned, and HAstV1 infected cells were visualized by indirect immunofluorescence (HAstV1 (red), nuclei (DAPI, blue). Scale bar 25 μm.</p><p><strong>C</strong>. Human intestinal organoids were incubated with media (mock) or infected with HAstV1. Organoids at 16 hpi were fixed and the presence of HAstV1 infected cells (green) were visualized by indirect immunofluorescence. Apical and basolateral membranes were immunostained for actin (magenta) and Laminin (red) respectively. Nuclei are stained with DAPI (blue). Scale bar is 20 μm.</p><p><strong>D.</strong> Quantification of C with the percentage of infected cells determined.</p><p><strong>E</strong>. Human intestinal organoids were infected with HAstV1. At indicated time post-infection the increase in viral copy number was determined by q-RT-PCR.</p><p><strong>F</strong>. Human intestinal organoids were incubated with media (mock) or infected with HAstV1. At 24 hpi the presence of IFNλ in the media was tested by ELISA. Dotted line indicates detection limit of the assay.</p><p><strong>G</strong>. Same as E but for the induced levels of either type I IFN (IFNβ1) or type III IFN (IFNλ).</p><p><strong>H</strong>. Human intestinal organoids were pre-treated for 24h with 2000IU/mL of IFNβ1 or 300ng/mL of IFNλ1-3. Interferons were maintained during the course of infection and the amount of HAstV1 copy numbers was assayed 24hpi by q-RT-PCR.</p><p>Data information: <strong>A-G</strong> Three biological replicates were performed for each experiment. Representative immunofluorescence images are shown. Error bars indicate the standard deviation. Statistics are from unpaired t-test.</p>
|
https://api.sourcedata.io/file.php?figure_id=41858
|
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"text": "Laminin",
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"text": "IFNβ1",
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"text": "type I IFN",
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"ext_dbs": "NCBI gene///NCBI gene",
"ext_ids": "282616///282617",
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"ext_tax_names": "Homo sapiens///Homo sapiens",
"mapping_source": "unknown",
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"ext_tax_names": "Homo sapiens///Homo sapiens///Homo sapiens///Homo sapiens",
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"text": "IFNλ",
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] |
||
10.15252/msb.20209833
|
Single-cell transcriptomics reveals immune response of intestinal cell types to viral infection
|
2021
|
Figure 3
|
<p><strong>Figure 3. Single-cell profiling and multiplex <em>in situ</em> RNA hybridization of human ileum-derived organoids.</strong></p><p><strong>A</strong>. UMAP of scRNA-Seq data from human ileum-derived organoids (n=16,682 cells); dots corresponding to cells are colored by the cell type.</p><p><strong>B</strong>. Dot plot of the top-three marker genes for each cell type. The dot size represents the percentage of cells expressing the gene; the color represents average expression across the cell type. Fig.S3B shows an extended list of the marker genes. All cell-type-defining marker genes are provided in Dataset EV2.</p><p><strong>C</strong>. Representative images showing multiplex <em>in situ</em> RNA hybridization of marker genes in an organoid. DAPI in blue.</p><p><strong>D-I</strong>. Visualizations for cell-type markers for the major intestinal cell types, showing single-cell expression intensities of the cell-type markers on the UMAP (left) and multiplex <em>in situ</em> RNA hybridization; red corresponds to viral RNA and a green corresponds to cell type markers (right).</p><p><strong>J-M.</strong> Percentage of stem cells (OLFM4) goblet cell (FGCBP), mature enterocytes (APOA4) and cells of the enterocyte lineage (FGCBP) from the total organoid cells population (black bar) in mock-treated organoids and after HAstV1 infection. The percentage of infected cells (red bars) and bystander cells (blue bars) within each cell type is shown. The data was obtained from multiplex in situ RNA hybridization in 2D organoids. n=10, mean ± SEM. Statistics show comparison of infected cells between mock-treated, 4h pi and 16h pi organoids. An Ordinary one-way ANOVA and Tukey's multiple comparisons test was used. n.s non significant, * p<0.05, ** p<0.01, *** p<0.001.</p>
|
https://api.sourcedata.io/file.php?figure_id=41862
|
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"mapping_status": "mapped",
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"original_type": "gene",
"role": "assayed",
"text": "APOA4",
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"uniprot_ids": [
"P06727"
]
},
{
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"text": "FGCBP",
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"ext_dbs": "NCBI gene",
"ext_ids": "10562",
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"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "10562",
"original_type": "gene",
"role": "assayed",
"text": "OLFM4",
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] |
||
10.15252/msb.20209833
|
Single-cell transcriptomics reveals immune response of intestinal cell types to viral infection
|
2021
|
Figure 5
|
<p><strong>Figure 5. Multiplex <em>in situ</em> RNA FISH visualizes immune signature in HastV1-infected and bystander cells.</strong></p><p><strong>A.</strong> One image per condition was used to show the multiplex in situ RNA FISH. A representative region of the images was chosen and cropped in order to have a zoom-in of a specific area for better visualization. Shown are the enterocyte lineage marker FABP6 (green), HAstV1 infection (white), and CCL2 gene expression (red) of mock-treated and infected organoids. DAPI is in blue. White arrows show co-localization of FABP6 and CCL2 with HAstV1 signal. Scale bar 100µm.</p><p><strong>B-D.</strong> Fluorescence intensity (arbitrary units) of innate immune marker expression in stem cells (OLFM4 positive), goblet cells (FCGBP positive), enterocyte lineage cells (FABP6 positive), and mature enterocytes (APOA4 positive). Bystander cells are in blue and infected cells in red. 10-90 percentile box plots, each dot represents one cell (mock N=2274, 4h N=3682, 16h N=1510. C. mock N=2693, 4h N=3706, 16h N=1921. D. mock N=2274, 4h N=3706, 16h N=1514.) . Blue statistics show a comparison of bystander cells between cell lineages and red statistics show the comparison of infected cells between cell lineages. Ordinary one-way ANOVA and Tukey's multiple comparisons test were used. Grey statistics show the comparison between infected and bystander cells within each cell lineage. Unpaired t-test with Welch's correction was used. n.s non significant, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.</p><h1 id="section-1"><br></h1><h1 id="extended-view-figure-legends">Extended View Figure Legends</h1>
|
https://api.sourcedata.io/file.php?figure_id=41866
|
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"mapping_status": "mapped",
"ncbi_gene_id": "6347",
"original_type": "gene",
"role": "assayed",
"text": "CCL2",
"type": "geneprod",
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]
},
{
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"text": "CCL2",
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"text": "FABP6",
"type": "geneprod",
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"P51161"
]
},
{
"ext_dbs": "NCBI gene",
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"text": "FABP6",
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] |
||
10.15252/embj.2021109783
|
Spt5 histone binding activity preserves chromatin during transcription by RNA polymerase II
|
2022
|
Figure 1
|
<p><strong>Figure 1</strong></p><p>Eukaryotic Spt5N binds histones H3-H4 via an evolutionarily conserved motif. (<strong>A</strong>) Domain structure of Spt5 / NusG proteins. The enlarged inset shows a conserved region of Spt5N from <em>Saccharomyces cerevisiae</em> (S.c.), <em>Schizosaccharomyces pombe</em> (S.p.), <em>Aspergillus nidulans</em> (A.n.), <em>Homo sapiens</em> (H.s.), <em>Xenopus laevis</em> (X.l.) and <em>Arabidopsis thaliana</em> (A.t.). Spt5N contains three invariant residues that were mutated in Spt5-3A (indicated by red asterisks), whilst residues with conservative substitutions are denoted by blue asterisks. (<strong>B</strong>) GST, GST-MCM2N (human) and GST-Spt5N (yeast) were mixed as indicated with yeast histones H3-H4, in the presence of glutathione agarose beads (see STAR method for further details). The input and pulldown samples were analysed by SDS-PAGE before staining with colloidal Coomassie blue. The asterisks denote truncations of GST-Spt5N. (<strong>C</strong>) Similar experiment to (B) but with histones H2A-H2B. (<strong>D</strong>) Similar experiment to (B) to compare histone binding by GST-MCM2N and GST-MCM2N-2A (with Y81A Y90A mutations of conserved residues). (<strong>E</strong>) Analogous experiment to (D), comparing histone binding by GST-Spt5 and GST-Spt5N-3A (with F180A E184A V187A mutations of conserved residues). (<strong>F</strong>) Budding yeast Spt5N and Spt5N-3A were expressed as fusions to Protein A in budding yeast cells. Cell extracts were treated with DNase as indicated, to release nucleosomal histone complexes from chromatin, before immunoprecipitation (IP) of Spt5N / Spt5N-3A on magnetic beads coated with IgG. The bound proteins were eluted and then analysed by mass spectrometry (the panel indicates the detected spectral counts). (<strong>G</strong>) Samples from an analogous experiment to that shown in (F) were analysed by immunoblotting of the indicated factors. The asterisk indicates a non-specific band in the immunoblot for histone H2B. (<strong>H</strong>). Budding yeast cells expressing fusions of Spt5N and Spt5N-3A to Protein A were grown in parallel with cells expressing the equivalent wild type and mutant versions of fission yeast Spt5N (Sp Spt5N and Sp Spt5N-3A - the latter contained the F146A, E150A and V153A mutations). See also Figure EV1.</p>
|
https://api.sourcedata.io/file.php?figure_id=45929
|
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"ext_tax_names": "Schizosaccharomyces pombe 972h-",
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"text": "Spt5",
"type": "geneprod",
"uniprot_ids": [
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||
10.15252/embj.2021109783
|
Spt5 histone binding activity preserves chromatin during transcription by RNA polymerase II
|
2022
|
Figure 2
|
<p><strong>Figure 2</strong></p><p>Mutation of the histone-binding motif of Spt5N is lethal, even in the presence of wild type Spt5. (<strong>A</strong>) Linearised versions of the indicated plasmids were transformed into <em>SPT5-9MYC</em> budding yeast cells and integrated at the <em>ura3</em> locus. For each transformation, four colonies were analysed by immunoblotting and DNA sequencing as shown. Colonies transformed with plasmid expressing Spt5-3A either lost the plasmid (1), contained the <em>SPT5-3A</em> DNA but failed to express Spt5-3A protein (2+4) or experienced a gene conversion and thus expressed wild type Spt5 from the integrated plasmid DNA (3). (<strong>B</strong>) Serial dilutions of yeast cells expressing the indicated fragments of wild type Spt5 or Spt5-3A from the <em>GAL1,10</em> promoter were grown on the indicated medium, together with a control of wild type cells. (<strong>C</strong>) Yeast cells were generated that expressed Protein A-tagged versions of wild type Spt5 (full length or the indicated truncations), or full length Spt5-3A, under the control of the <em>GAL1,10</em> promoter. Cells were grown in medium containing galactose and the ProteinA-tagged Spt5 proteins were isolated from cell extracts on magnetic beads coated with IgG. Association of the tagged Spt5 proteins with the elongating form of Pol II was monitored by immunoblotting, using antibodies specific to the indicated phosphorylation sites in the C-Terminal Domain repeats of the Rpb1 subunit of Pol II. (<strong>D</strong>) Control and <em>spt5-AID</em> strains were grown at 30°C in rich medium, before addition of auxin (0.5 mM 3-indoleacetic acid or IAA) for 1 hour. Spt5 protein was monitored in cell extracts by immunoblotting. (<strong>E</strong>) Serial dilutions of control cells or <em>spt5-AID</em> cells, expressing the indicated versions of Spt5 from the <em>GAL1,10</em> promoter, were grown on the indicated medium.</p>
|
https://api.sourcedata.io/file.php?figure_id=45931
|
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"P27692"
]
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"uniprot_ids": [
"A0A6V8S3R7"
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},
{
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]
},
{
"ext_dbs": "NCBI gene",
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"ext_tax_names": "Saccharomyces cerevisiae S288C",
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"mapping_status": "mapped",
"ncbi_gene_id": "854999",
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"role": "intervention",
"text": "Spt5",
"type": "geneprod",
"uniprot_ids": [
"P27692"
]
}
] |
||
10.15252/embj.2021109783
|
Spt5 histone binding activity preserves chromatin during transcription by RNA polymerase II
|
2022
|
Figure 3
|
<p><strong>Figure 3</strong></p><p>The histone-binding activity of Spt5N is dispensable for progression of the Pol II elongation complex through transcription units. (<strong>A</strong>) Experimental procedure, based on the yeast strains YCE281, YCE350 and YCE356. (<strong>B</strong>) Immunoblot analysis at the indicated times for the samples described in (A). (<strong>C</strong>) Cells from the end of the experiment described in (A) were fixed and processed for ChIP-Seq, using antibodies specific for phosphorylated Serine 5 in the C-Terminal Domain of the Pol II catalytic subunit Rpb1 (CTD-Ser5P). For each of the three indicated strains, the histograms represent the normalized read density (DNA sequence read count per million reads, or RPM), across a sample gene body. (<strong>D</strong>) Average of the normalised read density (RPM) for the CTD-Ser5P ChIP-Seq data from two independent experiments, for regions spanning 1kb upstream and downstream of the Transcription Start Site (TSS) of actively transcribed genes across the yeast genome (the analysis focussed on 821 actively transcribed genes, as described in Materials and Methods). (<strong>E</strong>) For each transcribed gene, an RNA Pol II Traveling ratio was calculated as indicated. (<strong>F</strong>) Log2 values of the calculated Pol II traveling ratios for each actively transcribed gene were arranged in order from low to high and then plotted against percentile. (<strong>G</strong>) Principal component analysis of two independent experiments, for the Pol II CTD-Ser5P ChIP-Seq signal across the yeast genome in the three indicated strains.</p>
|
https://api.sourcedata.io/file.php?figure_id=45933
|
[
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"ext_ids": "H9NB31",
"ext_tax_ids": "4932",
"ext_tax_names": "Saccharomyces cerevisiae",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Rpb1",
"type": "geneprod",
"uniprot_ids": [
"H9NB31"
]
}
] |
||
10.15252/embj.2021109783
|
Spt5 histone binding activity preserves chromatin during transcription by RNA polymerase II
|
2022
|
Figure 4
|
<sd-panel><p><strong>Figure 4</strong></p> <p>Spt5N histone binding activity preserves nucleosome integrity during transcription. (<strong>A</strong>) Samples from the experiment described in Figure 3A were also processed for MNase-Seq. For each of the three indicated strains, the histograms represent the normalized DNA sequence read count per million reads (RPM), across a sample gene body. (<strong>B</strong>) Average of the normalised read density (RPM) for the MNase-Seq data from two independent experiments, for regions spanning 1kb upstream and downstream of the Transcription Start Site (TSS) of actively transcribed genes across the yeast genome (as above, the analysis focussed on 821 active genes, as described in Materials and Methods). (<strong>C</strong>) Samples from the same experiment were further processed for ChIP-Seq using antibodies to histone H3K4me3. For each of the three indicated strains, the normalised read count per million reads is presented across a sample gene body. (<strong>D</strong>) Average of the normalised read density (RPM) for the H3K4me3 ChIP-Seq datasets from two independent experiments, for regions spanning 1kb upstream and downstream of the Transcription Start Site (TSS) of actively transcribed genes. (<strong>E</strong>) Principal component analysis of two independent experiments, for the H3K4me3 ChIP-Seq signals across the yeast genome in the three indicated strains. (<strong>F</strong>) Ratio of the H3K4me3 ChIP-Seq signal (TSS to +500 bp) in cells expressing Spt5-3A and cells expressing wild type Spt5. The boxes extend from the 25<sup>th</sup> to 75<sup>th</sup> percentiles whilst the 'whiskers' illustrate the minimum and maximal values and the lines within each box correspond to the median values. The data are expressed as a function of the gene expression level, based on the read density of the ChIP-Seq data for CTD-Ser5P (from TSS to TTS) in Figure 3, ordered from lowest to highest values in four quartiles (colour coded as indicated). The correlation between H3K4me3 in cells expressing Spt5-3A (<em>spt5-AID GAL-SPT5-3A)</em> and control cells (<em>spt5-AID GAL-SPT5</em>) is progressively worse from Q1 to Q4, for genes expressed at progressively higher levels.</p></sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=45935
|
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"ext_ids": "A0A6A5Q536",
"ext_tax_ids": "4932",
"ext_tax_names": "Saccharomyces cerevisiae",
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"mapping_status": "mapped",
"ncbi_gene_id": null,
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"role": "assayed",
"text": "histone H3",
"type": "geneprod",
"uniprot_ids": [
"A0A6A5Q536"
]
}
] |
||
26666269
|
10.15252/emmm.201505505
|
Endothelial cell-derived Angiopoietin-2 is a therapeutic target in treatment-naive and Bevacizumab-resistant glioblastoma
|
2015
|
Figure 1
|
<p>Figure 1: Endothelial Ang-2 upregulation correlates with WHO grading in human gliomas. ELISA displaying human Ang-2 (A) and Ang-1 (B) level in serum of healthy patients (Ang-2 n=4; Ang-1 n=21) or patients with low grade glioma (WHOII) (n=5), anaplastic astrocytoma (WHOIII) (n=7) or glioblastoma (WHOIV) (n=39) are shown. The TCGA database was accessed to obtain gene expression level for Ang-1, Ang-2, Tie2 and Tie1 in GBM in comparison to normal brain (C). Ang-2 expression and quantitative analysis of Ang-2 in healthy human brain (n=3), low grade diffuse glioma (n=14), anaplastic astrocytoma (n=12) or glioblastoma (n=11) is shown (D). Co-staining of Ang-2 and vWF (endothelial cells), αSMA (mural cells) and IBA1 (microglia) in different glioblastoma specimen. Normal brain tissue was used to assess Ang-2 staining specificity (E). Ang-2 predicts survival of glioblastoma patients (F). If not indicated differently, Kruskall-Wallis (Dunn´s post test) was applied, ** p < 0.05, *** p < 0.005; data are mean ± SEM</p>
|
https://api.sourcedata.io/file.php?figure_id=5251
|
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"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
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"uniprot_ids": [
"O15123"
]
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{
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"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O15123"
]
},
{
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"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
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"O15123"
]
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{
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"mapping_status": "mapped",
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"role": "assayed",
"text": "Ang-1",
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"text": "Ang-1",
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"B4DTQ9"
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"mapping_status": "mapped",
"ncbi_gene_id": "11601",
"original_type": "gene",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O35608",
"B9EHQ4"
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},
{
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"mapping_status": "mapped",
"ncbi_gene_id": "7010",
"original_type": "gene",
"role": "assayed",
"text": "Tie2",
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"Q02763",
"Q59HG2"
]
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{
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"role": "assayed",
"text": "Tie1",
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"uniprot_ids": [
"P35590",
"B4DTW8"
]
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{
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"uniprot_ids": [
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"ext_ids": "O15123",
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"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O15123"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P62736",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "αSMA",
"type": "geneprod",
"uniprot_ids": [
"P62736"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P55008",
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"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "IBA1",
"type": "geneprod",
"uniprot_ids": [
"P55008"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O15123",
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"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O15123"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O15123",
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"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O15123"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P04275",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "vWF",
"type": "geneprod",
"uniprot_ids": [
"P04275"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O15123",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O15123"
]
}
] |
|
26666269
|
10.15252/emmm.201505505
|
Endothelial cell-derived Angiopoietin-2 is a therapeutic target in treatment-naive and Bevacizumab-resistant glioblastoma
|
2015
|
Figure 2
|
<p>Figure 2: Endothelial Ang-2 expression reduced pericyte coverage and increased macrophage infiltration in experimental glioblastoma. GL261 cells were intracerebrally transplanted in wild type or Ang-2 gain-of-function mice (Ang-2 DT). Brain tumor sections were stained with antibodies against CD31 and desmin (A), and microvessel densities (MVD) and pericyte coverage were determined (WT n=14; Ang-2 DT n=6) (B). FACS analysis of infiltrating macrophages in Ang-2 DT normal brain (w/o tumor) compared to WT is shown (C, dot plot; D, quantification) (WT n=5; Ang-2 DT n=3). Immunohistochemistry staining for monocytes/macrophages (F4/80) and neutrophils (Ly6G; arrowheads) in brain tumors derived from WT or Ang-2 overexpressing mice (E), Quantitative analysis of C (F) (WT n=14; Ang-2 DT n=10). Kaplan-Meier survival curve of GL261 tumor-bearing WT and Ang-2 DT mice (WT n=17; Ang-2 DT n=14) (E). Statistical analyses were performed using an unpaired Student´s t-test (B, F), one-way Anova (Tukey post test) (D), Log-rank and Wilcoxon (G) * p < 0.05, ** p < 0.01; data are mean ± SEM</p>
|
https://api.sourcedata.io/file.php?figure_id=5252
|
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"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "11601",
"original_type": "gene",
"role": "intervention",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O35608",
"B9EHQ4"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "11601",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "11601",
"original_type": "gene",
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"text": "Ang-2",
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"uniprot_ids": [
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"B9EHQ4"
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},
{
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"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
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{
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},
{
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"text": "desmin",
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},
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{
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{
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"text": "F4/80",
"type": "geneprod",
"uniprot_ids": [
"Q61549"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P35461",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ly6G",
"type": "geneprod",
"uniprot_ids": [
"P35461"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "11601",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "11601",
"original_type": "gene",
"role": "intervention",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O35608",
"B9EHQ4"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "11601",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "11601",
"original_type": "gene",
"role": "intervention",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O35608",
"B9EHQ4"
]
}
] |
|
26666269
|
10.15252/emmm.201505505
|
Endothelial cell-derived Angiopoietin-2 is a therapeutic target in treatment-naive and Bevacizumab-resistant glioblastoma
|
2015
|
Figure 3
|
<p>Figure 3: Infiltrating leukocyte subsets in human glioma. Paraffin sections of healthy human brain (n=3), low grade glioma (n=12), anaplastic astrocytoma (n=7) and glioblastoma (n=9) were stained for IBA1 (macrophages/microglia), CD15 (granulocytes) and CD3 (T-cells) (A). Quantification of leukocyte subsets in human glioma specimens (B). For statistical analysis Kruskall-Wallis (Dunn´s post test) was applied.* p < 0.05, ** p < 0.01; data are mean ± SEM</p>
|
https://api.sourcedata.io/file.php?figure_id=5253
|
[
{
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"ext_ids": "P55008",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "IBA1",
"type": "geneprod",
"uniprot_ids": [
"P55008"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P07766",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD3",
"type": "geneprod",
"uniprot_ids": [
"P07766"
]
}
] |
|
26666269
|
10.15252/emmm.201505505
|
Endothelial cell-derived Angiopoietin-2 is a therapeutic target in treatment-naive and Bevacizumab-resistant glioblastoma
|
2015
|
Figure 4
|
<p>Figure 4: Dual anti-Ang-2 and anti-VEGF therapy acts synergistic on vascular normalization and macrophage infiltration in experimental GBM. Immunofluorescence staining of CD31 and desmin in GL261 glioblastoma after single and dual treatment with anti-Ang-2 (AMG386) and anti-VEGF (aflibercept) (A). Corresponding quantitative analysis of microvessel densities and pericytes numbers (B; control n=24; AMG386 n=10; aflibercept n=12; AMG386 + aflibercept n=13). Staining for mouse IgG (red) as a surrogate parameter for vascular leakage and CD31 (green) is shown in GL261 glioblastoma sections following different anti-angiogenic treatments (C) with quantification in (D; n=3). Quantification of necrotic areas in GL261 glioblastoma sections after anti-angiogenic treatment is shown as percent of whole tumor area (E; control, aflibercept, AMG386 + aflibercept n=4; AMG386 n=3). Analysis of Glut1 immunoreactivity in GL261 glioblastoma sections (F; n=3). Double-immunofluorescence stainings of macrophages (F4/80) and tumor vessels (vWF) in mouse GBM after treatment with AMG386, aflibercept or the combination of both are shown in (G). Quantitatve analysis of tumor infiltrating leukocytes (F4/80+, CD3+, Ly-6G+) following anti-angiogenic treatment is displayed in (H; control n=28; AMG386 n=12; aflibercept n=11; AMG386 + aflibercept n=12). Kaplan-Meier survival analysis of GL261 tumors grown in C57BL/6 mice following anti-angiogenic treatment (I; control n=38; AMG386 n=13; aflibercept n=12; AMG386 + aflibercept n=13). Double-immunofluorescence stainings with anti-F4/80 and anti-CD206 in brain tumor sections of mice treated with anti-Ang-2 (AMG386), anti-VEGF (aflibercept) or the combination of both are shown in (J). Corresponding quantitative analyses of tumor infiltrating F4/80+ cells and CD206+ cells, and the ratio of CD206+ vs. F4/80+ upon anti-angiogenic therapy are displayed in (K; control n=21; AMG386 n=13; aflibercept n=9; AMG386 + aflibercept n=4). Flow cytometry of tumor infiltrating macrophages following enzymatic dissociation of mouse GL261 brain tumors plus/minus anti-angiogenic therapy. Percent of CD206+ (L) and MHC class IIHI cells (M) among CD45+CD11b+GR1-F4/80+ cells are displayed (L,M; control n=4; AMG386 n=4; aflibercept n=5; AMG386 + aflibercept n=5). Statistical analyses were performed using one-way ANOVA and Tukey’s multiple comparison except for Log-rank and Wilcoxon (I) and Kruskall Wallis (B). * p < 0.05, ** p < 0.01, *** p < 0.005, # p < 0.005 of aflibercept + AMG386 vs aflibercept; data are mean ± SEM (A-K)</p>
|
https://api.sourcedata.io/file.php?figure_id=5254
|
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"text": "VEGF",
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]
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{
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"text": "vWF",
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]
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{
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{
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"P22646"
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"ext_dbs": "Uniprot",
"ext_ids": "P35461",
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"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
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"role": "assayed",
"text": "Ly-6G",
"type": "geneprod",
"uniprot_ids": [
"P35461"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "11601",
"ext_tax_ids": "10090",
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"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "11601",
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"text": "Ang-2",
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"O35608",
"B9EHQ4"
]
},
{
"ext_dbs": "NCBI gene",
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"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "22339",
"original_type": "gene",
"role": "intervention",
"text": "VEGF",
"type": "geneprod",
"uniprot_ids": [
"Q00731"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q61549",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
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"role": "assayed",
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]
},
{
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"ext_ids": "Q61549",
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"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "F4/80",
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"uniprot_ids": [
"Q61549"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q61549",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "F4/80",
"type": "geneprod",
"uniprot_ids": [
"Q61549"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q61830",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD206",
"type": "geneprod",
"uniprot_ids": [
"Q61830"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q61830",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD206",
"type": "geneprod",
"uniprot_ids": [
"Q61830"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q61830",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD206",
"type": "geneprod",
"uniprot_ids": [
"Q61830"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q61549",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "F4/80",
"type": "geneprod",
"uniprot_ids": [
"Q61549"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P24788",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD11b",
"type": "geneprod",
"uniprot_ids": [
"P24788"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P47791",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "GR1",
"type": "geneprod",
"uniprot_ids": [
"P47791"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q61830",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD206",
"type": "geneprod",
"uniprot_ids": [
"Q61830"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P06800",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD45",
"type": "geneprod",
"uniprot_ids": [
"P06800"
]
}
] |
|
26666269
|
10.15252/emmm.201505505
|
Endothelial cell-derived Angiopoietin-2 is a therapeutic target in treatment-naive and Bevacizumab-resistant glioblastoma
|
2015
|
Figure 5
|
<p>Figure 5: Anti-VEGF therapy led to decreased infiltration of CD68+ macrophages in human GBM. Anti-CD68 (A), anti-CSFR1 (B) and anti-CD206 (C) immunohistochemistry of patient samples derived from treatment-naive GBM (n=24), post-radiochemotherapy (S/RTx/CTx) (n=7) and post-radiochemotherapy + bevacizumab (S/RTx/CTx/Bev) (n=29) therapy is shown. Corresponding quantification of tumor infiltrating cells is displayed (anti-CD68, D), (anti-CSF1R, E) and (anti-CD206, F). Ratio of CD206+ vs. CD68+ cells (G). Amount of CD206+ perivascular cells per blood vessel (H). Double staining of Ang-2 (brown) and CD206 (red) of a S / RTx / CTx / Bev patient, arrowheads indicating perivascular CD206+ cells around Ang-2+ vessels (I). Arrows pointing on parenchymal CD206+ cells (I). CD206+ cells in relation to the percentage of hypoxic (CAIX positive) tumor area (J). Statistical analysis: Kruskall-Wallis (Dunn´s post test). * p < 0.05, ** p < 0.01; data are mean ± SEM</p>
|
https://api.sourcedata.io/file.php?figure_id=5255
|
[
{
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"ext_ids": "7422",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "7422",
"original_type": "gene",
"role": "intervention",
"text": "VEGF",
"type": "geneprod",
"uniprot_ids": [
"P15692",
"A0A0Y0IMM4",
"A2A2V4"
]
},
{
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"ext_ids": "P34810",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD68",
"type": "geneprod",
"uniprot_ids": [
"P34810"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P34810",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD68",
"type": "geneprod",
"uniprot_ids": [
"P34810"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P07333",
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"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CSFR1",
"type": "geneprod",
"uniprot_ids": [
"P07333"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P22897",
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"text": "CD206",
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"uniprot_ids": [
"P22897"
]
},
{
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"ext_ids": "P34810",
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"P34810"
]
},
{
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"ext_ids": "P34810",
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"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD68",
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"uniprot_ids": [
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]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P07333",
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"role": "assayed",
"text": "CSF1R",
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},
{
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},
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},
{
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"text": "Ang-2",
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{
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"ext_ids": "O15123",
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"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O15123"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P22897",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD206",
"type": "geneprod",
"uniprot_ids": [
"P22897"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P22897",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD206",
"type": "geneprod",
"uniprot_ids": [
"P22897"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P22897",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD206",
"type": "geneprod",
"uniprot_ids": [
"P22897"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q16790",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CAIX",
"type": "geneprod",
"uniprot_ids": [
"Q16790"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P22897",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD206",
"type": "geneprod",
"uniprot_ids": [
"P22897"
]
}
] |
|
26666269
|
10.15252/emmm.201505505
|
Endothelial cell-derived Angiopoietin-2 is a therapeutic target in treatment-naive and Bevacizumab-resistant glioblastoma
|
2015
|
Figure 6
|
<p>Figure 6: Bevacizumab therapy leads to reduced vessel density and increased Ang-2 expression. Immunohistochemistry stainings with antibodies directed against CD31 (A) and Ang-2 (B) in treatment-naive GBM, post-radiochemotherapy (S/RTx/CTx) and post-radiochemotherapy + bevacizumab (S/RTx/CTx/Bev) therapy are displayed. Quantitative analyses of microvessel densities and Ang-2+ vessels are shown in (C) and (D). The ratio of Ang-2+ vs. CD31+ vessels was determined in (E). Western Blot and corresponding quantification of CD31 and Ang-2 in control and aflibercept treated mice (F). CAIX staining of treatment-naive GBM and S / RTx / CTx / Bev patients (G). Analyses of hypoxic (CAIX+) area with regard to the whole tumor in human GBM (H). Tumor-infiltrating lymphocytes (CD8/CD3) in human patients (I). Kruskall-Wallis (Dunn´s post test). * p < 0.05, *** p < 0.005; treatment-naive GBM n=26, S/RTx/CTx n=7, S/RTx/CTx/Bev n=29; data are mean ± SEM</p>
|
https://api.sourcedata.io/file.php?figure_id=5256
|
[
{
"ext_dbs": "Uniprot",
"ext_ids": "O15123",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O15123"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O15123",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O15123"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P16284",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD31",
"type": "geneprod",
"uniprot_ids": [
"P16284"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O15123",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O15123"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O15123",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O15123"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P16284",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD31",
"type": "geneprod",
"uniprot_ids": [
"P16284"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O35608",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Ang-2",
"type": "geneprod",
"uniprot_ids": [
"O35608"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q08481",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD31",
"type": "geneprod",
"uniprot_ids": [
"Q08481"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q16790",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CAIX",
"type": "geneprod",
"uniprot_ids": [
"Q16790"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q16790",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CAIX",
"type": "geneprod",
"uniprot_ids": [
"Q16790"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P07766",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD3",
"type": "geneprod",
"uniprot_ids": [
"P07766"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P01732",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "CD8",
"type": "geneprod",
"uniprot_ids": [
"P01732"
]
}
] |
|
30573670
|
10.15252/embj.201899466
|
CRISPR/Cas9 searches for a protospacer adjacent motif by lateral diffusion
|
2018
|
Figure 3
|
<sd-panel> <p><strong>Figure 3. Multiple PAM influence on binding to a target site</strong></p> <ol type="A"> <li> <p>DNA and crRNA sequence used for experiments of a single target and multiple PAMs. DNA sequence is shown for the single-target single-PAM construct and schematic representation is shown for multiple-PAM containing constructs.</p> </li> <li> <p>FRET histogram of all binding events from the construct containing a single PAM next to the partial target. Y axis represents total number of counts.</p> </li> <li> <p>Example FRET traces from single-target multiple-PAM experiments. Top trace shows an example of a single-step binding event and a binding event that starts at a lower FRET state before transitioning to productive binding high-FRET state. Bottom trace shows an example of FRET fluctuations between the high-FRET state and lower FRET states.</p> </li> <li> <p>A histogram showing the percentage of dynamic. The percentage values were obtained from five experiments on five different days. Error bars represent standard error of the mean. The percentage of events showing dynamic binding behavior was calculated by dividing the number of binding events that showed such behavior by the total number of binding events.</p> </li> <li> <p>A scatter plot showing the time Cas9 spends in a lower FRET state before transitioning to a high-FRET state. The dwelltimes increase with the number of PAMs adjacent to the target. Such dwelltime could not be determined for a construct containing a single PAM. The values were obtained from five experiments on five different days. Error bars represent standard error of the mean.</p> </li> </ol> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=24324
|
[
{
"ext_dbs": "Uniprot",
"ext_ids": "A0A2W5CL48",
"ext_tax_ids": "",
"ext_tax_names": "",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Cas9",
"type": "geneprod",
"uniprot_ids": [
"A0A2W5CL48"
]
}
] |
|
30573670
|
10.15252/embj.201899466
|
CRISPR/Cas9 searches for a protospacer adjacent motif by lateral diffusion
|
2018
|
Figure 4
|
<sd-panel> <p><strong>Figure 4. Mechanism of lateral diffusion</strong></p> <ol type="A"> <li> <p>DNA sequences used in tandem target experiments. Complementary sequences to crRNA are marked in pink boxes and PAMs are marked in green boxes. Distances between the protospacers are indicated above the sequences. An illustration of Cas9 binding to each target site resulting in either high FRET (target H) or low FRET (target L) is shown on the right.</p> </li> <li> <p>Example fluorescence traces showing Cas9 transitioning between H and L targets with increasing target distances (right). FRET histograms constructed from events that show fluctuations showing two FRET peaks corresponding to binding to either target. High FRET peak stays the same in each histogram, but lower FRET peak moves to lower FRET values as the distance between targets increases.</p> </li> <li> <p>A scatter plot showing the probability of Cas9 transitioning between targets before dissociation. The values are averages from five measurements and error bars represent standard error of the mean. The transition probability (<em>p = T /</em> (<em>T+D</em>)) was calculated by summing up all the number of FRET transitions (<em>T</em>) and dividing <em>T</em> by the sum of <em>T</em> and the total number of dissociation events (<em>D</em>).</p> </li> <li> <p>A histogram showing all dwelltimes from single-target controls and tandem-target experiments. τ<sub>1</sub> is similar for single-target control and tandem target experiments. τ<sub>2</sub> only appears in tandem target experiments and is over 20-fold higher than τ<sub>1</sub> or τ<sub>st</sub>. The values represent the mean dwelltimes that were obtained by fitting dwelltime distributions by either single or double exponential decay function from five measurements. Error bars represent standard error of the mean.</p> </li> </ol> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=24325
|
[
{
"ext_dbs": "Uniprot",
"ext_ids": "A0A2W5CL48",
"ext_tax_ids": "",
"ext_tax_names": "",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Cas9",
"type": "geneprod",
"uniprot_ids": [
"A0A2W5CL48"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "A0A2W5CL48",
"ext_tax_ids": "",
"ext_tax_names": "",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Cas9",
"type": "geneprod",
"uniprot_ids": [
"A0A2W5CL48"
]
}
] |
|
10.1101/2020.03.24.005298
|
A unifying structural and functional model of the coronavirus replication organelle: tracking down RNA synthesis
|
2020
|
Fig. 6.
|
<sd-panel> <p>IEM detection of viral markers in MERS-CoV-infected cells.(A-G) Immunogold labeling of thawed cryo-sections of MERS-CoV-infected Huh7 cells (12 hpi) for the detection of the indicated viral proteins. (A-C) Structural proteins were detected on virions (black arrowheads) and, for the M and S proteins, also on Golgi cisterna. While regions containing DMS (white arrowheads) and CM labelled for the N protein (D) and nsp3 (G), the M and S protein were not detected in these areas. (H-I) Immunogold labeling of dsRNA in HPF-FS samples of MERS-CoV-infected Huh7 cells (13 hpi). The label accumulated on DMVs, which could be easily detected in this type of samples (grey arrowheads), while the regions with CM and DMSs, which appeared as dark areas among the DMV clusters, where devoid of dsRNA signal. G, Golgi complex; m, mitochondria. Scale bars, 250 nm.</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=33434
|
[
{
"ext_dbs": "Uniprot",
"ext_ids": "R9UNX5",
"ext_tax_ids": "1335626",
"ext_tax_names": "Middle East respiratory syndrome-related coronavirus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "M",
"type": "geneprod",
"uniprot_ids": [
"R9UNX5"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "R9UNX5",
"ext_tax_ids": "1335626",
"ext_tax_names": "Middle East respiratory syndrome-related coronavirus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "M",
"type": "geneprod",
"uniprot_ids": [
"R9UNX5"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "A0A141LLD8",
"ext_tax_ids": "1335626",
"ext_tax_names": "Middle East respiratory syndrome-related coronavirus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "N protein",
"type": "geneprod",
"uniprot_ids": [
"A0A141LLD8"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "A0A141LLB8",
"ext_tax_ids": "1335626",
"ext_tax_names": "Middle East respiratory syndrome-related coronavirus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "nsp3",
"type": "geneprod",
"uniprot_ids": [
"A0A141LLB8"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "W6A028",
"ext_tax_ids": "1335626",
"ext_tax_names": "Middle East respiratory syndrome-related coronavirus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "S protein",
"type": "geneprod",
"uniprot_ids": [
"W6A028"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "W6A028",
"ext_tax_ids": "1335626",
"ext_tax_names": "Middle East respiratory syndrome-related coronavirus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "S proteins",
"type": "geneprod",
"uniprot_ids": [
"W6A028"
]
}
] |
||
30530633
|
10.15252/embr.201846221
|
Mouse fetal intestinal organoids: new model to study epithelial maturation from suckling to weaning
|
2018
|
Figure 2
|
<sd-panel><p><strong>Figure 2</strong> - <strong>Small <sd-pretag id="sdPretag283779571sm" type="tissue" role="component">intestinal fetal</sd-pretag> organoids recapitulate suckling-to-weaning transition <em>in vitro</em></strong>.</p><p> <strong>A-G</strong> <sd-pretag id="sdPretag685066530sm" category="assay">Real</sd-pretag>-<sd-pretag id="sdPretag1544831807sm" category="assay">time qPCR</sd-pretag> analysis of fetal organoids cultured for one month showing decrease in relative expression of indicated neonatal markers <strong>A</strong> <sd-pretag id="sdPretag961315792sm" type="geneprod" role="assayed"><em>Ass1</em></sd-pretag>, <strong>B</strong> <sd-pretag id="sdPretag110127558sm" type="geneprod" role="assayed"><em>Blimp-1</em></sd-pretag>, <strong>C</strong> <em>FcRn</em> and <strong>D</strong> <sd-pretag id="sdPretag787166546sm" type="geneprod" role="assayed"><em>Lct</em></sd-pretag> and increase of the adult markers <strong>E</strong> <em>Sis</em>, <strong>F</strong> <em>Treh</em> and <strong>G</strong> <sd-pretag id="sdPretag2037167415sm" type="geneprod" role="intervention"><em>Arg2</em></sd-pretag> (n=3 individual wells from single organoid culture (see material and methods); experiment was repeated in four to eight independent organoid cultures with similar results). <strong>H-L.</strong> Enzyme <sd-pretag id="sdPretag8208300sm" category="assay">activity assay</sd-pretag> of fetal organoids for <strong>H</strong> Lactase, <strong>I</strong> sucrase, <strong>J</strong> maltase, <strong>K</strong> trehalase and <strong>L</strong> arginase. Activity is given in µM <sd-pretag id="sdPretag993971913sm" type="molecule" role="component">glucose</sd-pretag>/µg <sd-pretag id="sdPretag1088299090sm" type="geneprod" role="intervention">protein·min<sup>-1</sup></sd-pretag> (experiment was generated from single organoid culture (see material and methods) and repeated in 3 independent organoid cultures with similar results).</p><p> Data information: Data are presented as mean ± SEM. ND not detected, *p<0.05, **p<0.01, ***p<0.001, NS not significant, relative to expression or activity level at day 3 (one-way ANOVA).</p></sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=23969
|
[
{
"ext_dbs": "NCBI gene",
"ext_ids": "11847",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "11847",
"original_type": "gene",
"role": "assayed",
"text": "Arg2",
"type": "geneprod",
"uniprot_ids": [
"O08691"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "11898",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "11898",
"original_type": "gene",
"role": "assayed",
"text": "Ass1",
"type": "geneprod",
"uniprot_ids": [
"P16460",
"Q3UJ34"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "14132",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "14132",
"original_type": "gene",
"role": "assayed",
"text": "FcRn",
"type": "geneprod",
"uniprot_ids": [
"Q61559",
"Q6PKB0"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "226413",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "226413",
"original_type": "gene",
"role": "assayed",
"text": "Lct",
"type": "geneprod",
"uniprot_ids": [
"F8VPT3"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "12142",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "12142",
"original_type": "gene",
"role": "assayed",
"text": "Blimp-1",
"type": "geneprod",
"uniprot_ids": [
"Q60636"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "69983",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "69983",
"original_type": "gene",
"role": "assayed",
"text": "Sis",
"type": "geneprod",
"uniprot_ids": [
"F8VQM5"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "58866",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "58866",
"original_type": "gene",
"role": "assayed",
"text": "Treh",
"type": "geneprod",
"uniprot_ids": [
"Q9JLT2",
"E9PYP7"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O08691",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "arginase",
"type": "geneprod",
"uniprot_ids": [
"O08691"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P70699",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "maltase",
"type": "geneprod",
"uniprot_ids": [
"P70699"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "F8VPT3",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "Lactase",
"type": "geneprod",
"uniprot_ids": [
"F8VPT3"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "F8VQM5",
"ext_tax_ids": "10090",
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"ext_ids": "Q9JLT2",
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"ext_tax_names": "Mus musculus",
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] |
|
30530633
|
10.15252/embr.201846221
|
Mouse fetal intestinal organoids: new model to study epithelial maturation from suckling to weaning
|
2018
|
Figure 3
|
<sd-panel><p><strong>Figure 3</strong> - <strong>Fetal organoids resemble adult organoids after one month in culture</strong>.</p><p> <strong>A-G</strong> Relative expression detected by <sd-pretag id="sdPretag1421187240sm" category="assay">Real</sd-pretag>-<sd-pretag id="sdPretag897429407sm" category="assay">time qPCR</sd-pretag> in fetal organoids ( ) and adult organoids ( ). Neonatal markers <strong>A</strong> <sd-pretag id="sdPretag1095698709sm" type="geneprod" role="assayed"><em>Ass1</em></sd-pretag>, <strong>B</strong> <sd-pretag id="sdPretag1042392998sm" type="geneprod" role="assayed"><em>Blimp-1</em></sd-pretag>, <strong>C</strong> <em>FcRn</em> and <strong>D</strong> <sd-pretag id="sdPretag1469947426sm" type="geneprod" role="assayed"><em>Lct</em></sd-pretag> in fetal organoids cultured for one month compare to levels adult organoids. Mature markers <strong>E</strong> <em>Sis</em>, <strong>F</strong> <em>Treh</em> and <strong>G</strong> <sd-pretag id="sdPretag574938972sm" type="geneprod" role="assayed"><em>Arg2</em></sd-pretag> increase in fetal organoids to adult levels (n=3 individual wells from single organoid culture (see material and methods); experiment was repeated in four independent organoid cultures with similar results). <strong>H-L</strong> <sd-pretag id="sdPretag757665261sm" category="assay">Enzyme activity assay</sd-pretag> of fetal ( ) and adult ( ) organoids. <strong>H</strong> Lactase activity is higher in fetal organoids decreasing to adult levels at day 28, while <strong>I</strong> sucrase, <strong>J</strong> maltase, <strong>K</strong> trehalase and <strong>L</strong> <sd-pretag id="sdPretag1071632025sm" type="geneprod" role="assayed">arginase</sd-pretag> increase to adult levels. Activity is given in µM <sd-pretag id="sdPretag101919257sm" type="molecule" role="component">glucose</sd-pretag>/µg <sd-pretag id="sdPretag1940678281sm" type="geneprod" role="intervention">protein·min<sup>-1</sup></sd-pretag> (n=3 independent organoid cultures).</p><p> Data information: Data are presented as mean ± SEM. ND not detected, *p<0.05, **p<0.01, ***p<0.001, NS not significant, in A-G between fetal and adult organoids and in H-L relative to adult levels (t-test).</p></sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=23971
|
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"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "69983",
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{
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"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
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"Q9JLT2",
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"ext_tax_names": "Mus musculus",
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}
] |
|
30530633
|
10.15252/embr.201846221
|
Mouse fetal intestinal organoids: new model to study epithelial maturation from suckling to weaning
|
2018
|
Figure 4
|
<sd-panel><p><strong>Figure 4</strong> - <strong>Dexamethasone accelerates maturation of fetal organoids.</strong></p><p> <strong>A, B, E, F</strong> Maturation is accelerated in <sd-pretag id="sdPretag749573958sm" type="molecule" role="component">dexamethasone</sd-pretag> treated fetal organoids ( ) compared to control ( ), as revealed by <sd-pretag id="sdPretag614581570sm" category="assay">real</sd-pretag>-<sd-pretag id="sdPretag41993176sm" category="assay">time qPCR</sd-pretag>. Decrease in relative expression of neonatal marker <strong>A</strong> <sd-pretag id="sdPretag378794242sm" type="geneprod" role="assayed"><em>Blimp-1</em></sd-pretag> and increase in expression of adult marker <strong>B</strong> <em>Sis</em> occur earlier in <sd-pretag id="sdPretag1574965629sm" type="molecule" role="intervention">dexamethasone</sd-pretag> treated organoids, but expression of adult marker <strong>E</strong> <em>Treh</em> and <strong>F</strong> <em>Arg2</em> remains unchanged (n=3 individual wells from single organoid culture (see material and methods); experiment was repeated in four independent organoid cultures with similar results).</p><p> <strong>C,D,G,H</strong> <sd-pretag id="sdPretag520759737sm" type="molecule" role="component">Dexamethasone</sd-pretag> treatment accelerates increase in activity of <strong>E</strong> sucrase, <strong>F</strong> maltase, <strong>G</strong> trehalase and <strong>H</strong> arginase reaching adult organoid level ( ) at day 13 of the culture. Activity is given in µM <sd-pretag id="sdPretag284727737sm" type="molecule" role="component">glucose</sd-pretag>/µg <sd-pretag id="sdPretag398315401sm" type="geneprod" role="intervention">protein·min<sup>-1</sup></sd-pretag> (n = 3 independent organoid cultures).</p><p> Data information: Data are presented as mean ± SEM. ND not detected, *p<0.05, ***p<0.001, NS not significant, between control and <sd-pretag id="sdPretag473019715sm" type="molecule" role="intervention">dexamethasone</sd-pretag> treated organoids (two-way ANOVA). Expression values of control fetal organoids and control adult organoids in B, E and F are the same as Figure 3E-G and enzyme activity levels of fetal organoids in C, D, G and H are the same as 3I-L.</p></sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=23973
|
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]
},
{
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"mapping_status": "mapped",
"ncbi_gene_id": "69983",
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"text": "Sis",
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"uniprot_ids": [
"F8VQM5"
]
},
{
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"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "maltase",
"type": "geneprod",
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]
},
{
"ext_dbs": "Uniprot",
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]
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"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
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"role": "assayed",
"text": "Treh",
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"E9PYP7"
]
},
{
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"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "arginase",
"type": "geneprod",
"uniprot_ids": [
"O08691"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q9JLT2",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
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"text": "trehalase",
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"uniprot_ids": [
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]
}
] |
|
30530633
|
10.15252/embr.201846221
|
Mouse fetal intestinal organoids: new model to study epithelial maturation from suckling to weaning
|
2018
|
Figure 5
|
<sd-panel><p><strong>Figure 5</strong> - <strong><sd-pretag id="sdPretag1949810976sm" type="cell" role="component">Spheroids</sd-pretag> and <sd-pretag id="sdPretag779744354sm" type="tissue" role="component">organoids</sd-pretag> show the same protein expression pattern, independently of culture stage/passage.</strong></p><p> <strong>A</strong> <sd-pretag id="sdPretag85525050sm" category="assay">Microscopic</sd-pretag> images of fetal organoids at day 3, 13, 20 and 30 of culture. Scale bars: 500 µm.</p><p> <strong>B</strong> Percentage of fetal <sd-pretag id="sdPretag30024445sm" type="cell" role="component">spheroids</sd-pretag> decreases during culture (n=3 individual wells from single organoid culture (see material and methods); experiment was repeated in ten independent organoid cultures with similar results).</p><p> <strong>C-E</strong> <sd-pretag id="sdPretag1379799018sm" category="assay">Immmunohistochemistry</sd-pretag> of fetal organoids for <strong>C</strong> <sd-pretag id="sdPretag1573020100sm" type="geneprod" role="component">Ass1</sd-pretag> <strong>D</strong> Sis and <strong>E</strong> <sd-pretag id="sdPretag1903932062sm" type="geneprod" role="assayed">Arg2</sd-pretag>. Quantification of 3 slides per staining, 7 to 10 organoids / <sd-pretag id="sdPretag1553298788sm" type="cell" role="component">spheroids</sd-pretag> per slide. Scale bars: 50µm.</p><p> Data information: In B, data are presented as mean ± SEM. ***p<0.001, relative to number of <sd-pretag id="sdPretag1520371783sm" type="cell" role="component">spheroids</sd-pretag> at day 3 (one-way ANOVA).</p><p> <strong>Expanded View Figure Legends</strong></p></sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=23975
|
[
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"ncbi_gene_id": null,
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"text": "Arg2",
"type": "geneprod",
"uniprot_ids": [
"O08691"
]
},
{
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"ext_tax_names": "Mus musculus",
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{
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"text": "Sis",
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"uniprot_ids": [
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]
}
] |
|
10.15252/embr.202050145
|
Carbonic anhydrase 7 bundles filamentous actin and regulates dendritic spine morphology and density
|
2021
|
Figure 1
|
<sd-panel> <p><strong>Figure 1. Subcellular localization of CA7 and CA2 in fibroblasts.</strong></p> <p>(A) NIH3T3 fibroblasts co-expressing DsRed-CA7 and EGFP-CA2 (<em>n</em> = 4 independent replicates).</p> <p>(B-E) Co-localization of EGFP and the two CA isoforms with filamentous actin studied in fibroblasts expressing (B) EGFP, (C) EGFP-CA2, or (D, E) EGFP-CA7 and stained with phalloidin-594 to visualize F-actin (<em>n</em> = 4, 10 and 8 independent replicates, respectively). A magnification of the area marked with the yellow rectangle in (C) and (D) shows the localization of EGFP-CA2 and EGFP-CA7 compared to phalloidin-594. (E) EGFP-CA7 caused a prominent overexpression phenotype with thick and curvy cytosolic actin bundles (arrow) and plasmalemmal protrusions (arrowhead).</p> <p>(F-H) The normalized fluorescence emission intensity profiles for F-actin (red line) and (F) EGFP, (G) EGFP-CA2, or (H) EGFP-CA7 (black line).</p> <p>Data information: For the plots, pixel intensities were measured through the cross-section of the cell indicated by the yellow line in panels B-D. Scale bar in (A-E) 20 µm.</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=39022
|
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"text": "CA2",
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"text": "CA2",
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{
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"ext_tax_names": "Mus musculus",
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"mapping_status": "mapped",
"ncbi_gene_id": null,
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"text": "CA2",
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]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q9ERQ8",
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"ext_tax_names": "Mus musculus",
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"mapping_status": "mapped",
"ncbi_gene_id": null,
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"role": "assayed",
"text": "CA7",
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"uniprot_ids": [
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]
}
] |
||
10.15252/embr.202050145
|
Carbonic anhydrase 7 bundles filamentous actin and regulates dendritic spine morphology and density
|
2021
|
Figure 2
|
<sd-panel> <p><strong>Figure 2. CA7 binds to filamentous actin and increases actin bundling.</strong></p> <p>(A) Actin co-sedimentation assay shows that CA7 binds to F-actin. The binding is enhanced at more acidic pH (6.5 vs. 7.4) <em>n</em> = 3 independent replicates at each actin concentration, two-way ANOVA, P < 0.001.</p> <p>(B) Fluorescence time‐lapse images of F-actin bundling in an <em>in vitro</em> bundling assay. A mixture of unlabeled and Rhodamine labelled non-muscle actin was polymerized in the absence (PBS control, upper panel) or presence of mCA7 (lower panel). Numbers in images indicate the time after the onset of the experiment (0, 5 and 23 min). Intensity based Fire-coloring (Fiji) was used to visualize intensity changes. Scale bar 10 µm.</p> <p>(C) Quantification of the mean increase in filament length (<em>n</em> = 10 filaments at each time point) and the mean relative fluorescence intensity values of cross-sections for individual filaments /bundles (<em>n</em> = 30 - 31) in the absence and presence of mCA7 (1.12 µM). The data were analyzed using a general mixed model with time as a within-unit factor and the presence of CA7 as a between-unit factor. <em>n</em> = 3 independent repetitions, experiment repeats were included as a covariate and were non-significant.</p> <p>(D) Kymographs showing different time points in the line of interest (line width 1 µm) from the experiments analyzed in (C). Kymographs were generated with Fiji Multi Kymograph function. Total time is 159 frames = 26.5 minutes. Scale bar 5 μm (full height is 20 μm). F-actin bundles can be detected as clear lines in kymographs.</p> <p>(E) Fluorescence time‐lapse images of F-actin bundling in an <em>in vitro</em> bundling assay. A mixture of unlabeled and Rhodamine labelled non-muscle actin was polymerized in the presence of mCA7 (0.11 µM). Numbers above images indicate the time after the onset of the experiment (22 min -23 min 20 sec). Intensity based Fire-coloring (Fiji) was used to visualize intensity changes. White arrows highlight the bundling filaments. Scale bar 5 μm. This is a representative video from 8 similar experiments.</p> <p>Data information: Data are presented as mean ± SEM in (A) and mean ± SD in (C).</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=39024
|
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}
] |
||
10.15252/embr.202050145
|
Carbonic anhydrase 7 bundles filamentous actin and regulates dendritic spine morphology and density
|
2021
|
Figure 3
|
<sd-panel> <p><strong>Figure 3. CA7 overexpressing NIH3T3 are resistant to latrunculin B treatment.</strong></p> <p>(A,B) NIH3T3 cells transfected with DsRed (A) or DsRed-CA7 (B) were incubated in growth medium with 5 µM Latrunculin B for 0, 2, 5, 10, or 30 minutes, or in an equal amount of DMSO for 60 minutes. Analyses of experiments show that in cells transfected with DsRed-CA7 F-actin structures collapse more slowly (0 min: 83% "normal"; 2 min: 80%, p = 0.44; 5 min: 74%, p = 0.39; 10 min: 27%, p = 0.08; 30 min: 16%, p = 0.02; DMSO: 82%, p = 0.99; tested against 0 min with two-way ANOVA, Dunnett's multiple comparison test) than in the DsRed-transfected ones (0 min: 89% "normal"; 2 min: 48%, p = 0.14; 5 min: 31%, p = 0.04; 10 min: 2%, p = 0.001; 30 min: 0.7%, p = 0.001; DMSO: 88%, p = 0.8; tested against 0 min with two-way ANOVA, Dunnett's multiple comparison test) . For the analysis, cells were categorized to three groups as "normal", "some shape/F-actin left" and "round". The upper panel shows example images of the cells in all three categories for (A) DsRed- or (B) DsRed-CA7 transfected cells (actin visualized with Phalloidin-488).</p> <p>Data information: A hundred cells per each time point from each experiment (n = 3) were counted and categorized. Scale bar 50 µm. Data are presented as mean ± SEM.</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=39026
|
[
{
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"ncbi_gene_id": "12354",
"original_type": "gene",
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"text": "CA7",
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"Q9ERQ8",
"G3XA26",
"Q3UND9"
]
},
{
"ext_dbs": "NCBI gene",
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"ext_tax_names": "Mus musculus",
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"ncbi_gene_id": "12354",
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"Q9ERQ8",
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]
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{
"ext_dbs": "NCBI gene",
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"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
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"mapping_status": "mapped",
"ncbi_gene_id": "12354",
"original_type": "gene",
"role": "intervention",
"text": "CA7",
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"uniprot_ids": [
"Q9ERQ8",
"G3XA26",
"Q3UND9"
]
}
] |
||
10.15252/embr.202050145
|
Carbonic anhydrase 7 bundles filamentous actin and regulates dendritic spine morphology and density
|
2021
|
Figure 5
|
<sd-panel> <p><strong>Figure 5. Subcellular localization of the chimeric CA7 constructs in fibroblasts.</strong></p> <p>(A-C) NIH3T3 fibroblasts expressing EGFP-CA7-mutant1 (A), EGFP-CA7-mutant2 (B), and EGFP-CA7-mutant3 (C). F-actin is visualized with Phalloidin-594. In the right-most panel of (A-C) are the normalized fluorescence intensity profiles of the mutated CA7 EGFP signal (black) and actin (red) and the yellow line in left-most panels indicates the cross-section from which the pixel intensities were measured. Scale bars 20 µm.</p> <p>(D) Analysis of the mutated CA7 and F-actin co-localization in cultured fibroblasts. Scatterplots of fluorescent intensities per pixel (EGFP vs. Phalloidin-594) along a cross section through a representative cell. Pearson's correlation coefficient (r) for the analyzed cell is given in each panel.</p> <p>(E) Pearson's correlation coefficient values calculated for the depicted constructs and compared to CA7. F-actin had a strong positive correlation coefficient with EGFP-CA7 (r = 0.91 ± 0.02, <em>n</em> = 26 cells). Neither EGFP alone (r = 0.01 ± 0.03, <em>n</em> = 56) nor EGFP-CA2 (r = 0.09 ± 0.03, <em>n</em> = 53) co-localized with F-actin (<em>P</em> < 0.0001 for both constructs, when compared to CA7). From the five mutated CA7 constructs, EGFP-CA7-mutant1 (r= 0.51 ± 0.04, <em>P</em> < 0.0001, <em>n</em> = 25) and EGFP-CA7-mutant3 (r = 0.72 ± 0.03, <em>P</em> = 0.05, <em>n</em> = 21) co-localized less with F-actin actin when compared to CA7. The co-localization of the other four mutated CA7 constructs, EGFP-CA7-mutant2 (r = 0.92 ± 0.02, <em>P</em> = 0.99, <em>n</em> = 18), EGFP-CA7-R223E (r = 0.86 ± 0.02, <em>P</em> = 0.99, <em>n</em> = 24) and EGFP-CA7-H96/98C (r = 0.79 ± 0.04, <em>P</em> = 0.14, <em>n</em> = 18) did not differ significantly from that of CA7. Data are shown as mean ± SEM. Data did not pass Shapiro-Wilk test for normality, statistical comparison against CA7 was done with Kruskall-Wallis test corrected for multiple comparisons.</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=39030
|
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"text": "CA7",
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"uniprot_ids": [
"Q9ERQ8",
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},
{
"ext_dbs": "NCBI gene",
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"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
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"text": "CA7",
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] |
||
10.15252/embr.202050145
|
Carbonic anhydrase 7 bundles filamentous actin and regulates dendritic spine morphology and density
|
2021
|
Figure 6
|
<sd-panel> <p><strong>Figure 6. Localization of the overexpressed CA2 and CA7 in neurons.</strong></p> <p>(A) Isoform-specific subcellular localization shown in cultured hippocampal neurons (DIV14) co-expressing DsRed-CA7 (<em>left</em>) and EGFP-CA2 (<em>middle</em>).</p> <p>(B,C) Representative confocal images of precocious <em>in vivo</em> expression of (B) EGFP-CA2 and (C) EGFP-CA7 in P40 mouse cortical layer 2/3 pyramidal neurons. Neurons were transfected at E14.5 with EGFP-CA2 or EGFP-CA7 using <em>in utero</em> electroporation and images were taken from fixed slices. Right panels in (B) and (C) show higher magnification of the primary apical dendrite marked with a box.</p> <p>(D) The expression of EGFP-CA7 disrupted the normal spine morphology and induced the formation of thick, filopodia-like protrusions.</p> <p>Data information: <em>n</em> = 5 independent repeats for cultured neurons and two animals/construct <em>in vivo</em>. Scale bar in (A) 5 µm; (B and C): 5 µm, insets in (B, C) and panel D: 25 µm.</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=39032
|
[
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"ext_tax_names": "Rattus norvegicus",
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"text": "CA2",
"type": "geneprod",
"uniprot_ids": [
"P27139"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "B2RZ61",
"ext_tax_ids": "10116",
"ext_tax_names": "Rattus norvegicus",
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{
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{
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},
{
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"ncbi_gene_id": null,
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"text": "CA7",
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"uniprot_ids": [
"Q9ERQ8"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q9ERQ8",
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"ext_tax_names": "Mus musculus",
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"ncbi_gene_id": null,
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"role": "assayed",
"text": "CA7",
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"uniprot_ids": [
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}
] |
||
10.15252/embr.202050145
|
Carbonic anhydrase 7 bundles filamentous actin and regulates dendritic spine morphology and density
|
2021
|
Figure 7
|
<sd-panel> <p><strong>Figure 7. Layer 2/3 cortical pyramidal neurons in CA7 KO mice have high dendritic spine density and smaller spines but mEPSC frequency and amplitude are not affected</strong></p> <p>(A) Comparison of mEPSCs in cortical layer 2/3 pyramidal neurons from P30 - P40 WT and CA7 KO mice. Sample traces of mEPSC recordings from WT and CA7 KO neurons, low-pas filtered at 1 kHz (left). The data are summarized in the bar diagrams (right). mEPSC frequency (<em>P</em> = 0.63) and amplitude (<em>P</em> = 0.90) were not significantly different between WT and CA7 KO and neurons (<em>n</em> = 7 and 5 neurons, respectively, Student's independent samples <em>t</em>-test).</p> <p>(B) Representative confocal images of apical dendrites from Lucifer Yellow injected cortical layer 2/3 pyramidal neurons from WT and CA7 KO mice. The dendritic spine density and spine head size were examined in fixed slice preparations from P34 - P37 mice. Scale bar 2 µm.</p> <p>(C) Summary of the spine density analysis done from the Lucifer Yellow injected neurons. Spine density was a higher in CA7 KO neurons both in apical and basal dendrites (n = 28 neurons for both) compared to WT (n= 29 neurons for apical and n=30 for basal dendrite analysis) (<em>P</em> = 0.000002 for apical dendrites, analyzed with Mann-Whitney test, and <em>P</em> = 6,8 x 10<sup>-8</sup> for basal dendrites, Student's <em>t</em>-test with Welch-correction.) A total of 8279 spines were analyzed from four CA7 KO mice and 8730 spines from two WT control mice.</p> <p>(D) The spine head width distribution7differed significantly between the genotypes (n= 467 spines from WT and n = 421 spines from CA7 KO animals, 15 neurons analyzed from both genotypes, Wilcoxon rank sum test with continuity correction, W = 134540, <em>P</em> < 0.001).</p> <p>Data information: Data in (A) and (C) are given as mean + SEM.</p> <p>Expanded View Figure Legends</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=39033
|
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"mapping_status": "mapped",
"ncbi_gene_id": "12354",
"original_type": "gene",
"role": "intervention",
"text": "CA7",
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"Q3UND9"
]
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"ext_tax_names": "Mus musculus",
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"text": "CA7",
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"ext_dbs": "NCBI gene",
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"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
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"ncbi_gene_id": "12354",
"original_type": "gene",
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"Q9ERQ8",
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}
] |
||
27110680
|
10.1038/nchembio.1230
|
Chemical modulation of chaperone-mediated autophagy by retinoic acid derivatives
|
2013
|
figf1
|
<p>(<b>a</b>) Knockdown of <named-entity id="named-entity-253">RARα</named-entity> in NIH3T3 mouse fibroblasts was conducted using two different shRNAs, sh1 and sh2, compared to control (Ctr). Left, representative immunoblot. Actin is shown as loading control and full-length blots are shown in <sir rid="S1" refobjid="nchembio.1230-S1">Supplementary Figure 21</sir>. Right, amounts of <named-entity id="named-entity-254">RARα</named-entity> in control and knockdown cells determined by densitometric quantification of immunoblots represented by the one shown on the left. Values are normalized for actin and expressed as multiples of control (None) values; <i>n</i> = 3. (<b>b</b>) Rates of degradation of long-lived proteins in control and <named-entity id="named-entity-255">RARα</named-entity>(−) cells maintained in the presence or absence of serum for 12 h. Values are expressed as percentage of proteolysis; <i>n</i> = 3. (<b>c</b>,<b>d</b>) Percentage of degradation due to lysosomes (<b>c</b>) and macroautophagy (<b>d</b>) in cells assayed as in <b>b</b> but treated with inhibitors of lysosomal proteolysis (<b>c</b>) or with <named-entity id="named-entity-256">3-methyladenine</named-entity> (<named-entity id="named-entity-257">3MA</named-entity>) to block macroautophagy (<b>d</b>). Values are expressed as percentage of total protein degradation sensitive to the lysosomal inhibitors; <i>n</i> = 3. In all panels, all values are mean ± s.e.m., and differences with control are significant for *<i>P</i> 0.05.</p>
|
https://api.sourcedata.io/file.php?figure_id=3208
|
[
{
"ext_dbs": "NCBI gene",
"ext_ids": "19401",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "19401",
"original_type": "gene",
"role": "intervention",
"text": "RARα",
"type": "geneprod",
"uniprot_ids": [
"P11416",
"Q3U3R3",
"Q3U5E7"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P11416",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "RARα",
"type": "geneprod",
"uniprot_ids": [
"P11416"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "19401",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "19401",
"original_type": "gene",
"role": "intervention",
"text": "RARα",
"type": "geneprod",
"uniprot_ids": [
"P11416",
"Q3U3R3",
"Q3U5E7"
]
}
] |
|
27110680
|
10.1038/nchembio.1230
|
Chemical modulation of chaperone-mediated autophagy by retinoic acid derivatives
|
2013
|
figf2
|
<p>(<b>a</b>) Immunoblot for <named-entity id="named-entity-259">LC3-II</named-entity> in control mouse fibroblasts (Ctr) or those knocked down for <named-entity id="named-entity-260">RARα</named-entity> (<named-entity id="named-entity-261">RARα</named-entity>(−)) maintained in the presence or absence of serum for the indicated times. Where indicated, protease inhibitors (PI) against lysosomal proteolysis were added. Actin is shown as a loading control. 'I' and 'II' designate different forms of LC3, as described in the text. (<b>b</b>) Amounts of <named-entity id="named-entity-262">LC3-II</named-entity> determined by densitometric quantification of immunoblots. Values are expressed as fold change in value relative to serum-supplemented control cells; <i>n</i> = 4. (<b>c</b>) Ratio of the amounts of <named-entity id="named-entity-263">LC3-II</named-entity> in cells treated with protease inhibitors compared to untreated cells (None). Values are expressed as fold change in value relative to untreated cells; <i>n</i> = 4. (<b>d</b>,<b>e</b>) Autophagic flux in the same cells as in <b>c</b> expressing mCherry-GFP–<named-entity id="named-entity-264">LC3-II</named-entity> and maintained in the presence or absence of serum. Shown in <b>d</b> are representative merged channel images; scale bars, 2 μm. Arrows indicate autolysosomes (red). Shown in <b>e</b> are the quantification of the number of autophagosomes (mCherry and GFP positive) per cell (left) and percentage of autolysosomes (mCherry positive and GFP negative; right) in >50 cells in at least four different fields. AV, autophagic vacuoles. (<b>f</b>) Control and <named-entity id="named-entity-265">RARα</named-entity>(−) cells were transfected with the KFERQ-mCherry1 photoactivatable reporter, and after photoactivation they were maintained in medium with or without serum. Left, representative images. Graph shows quantification of the number of puncta per cell in >50 cells using the CMA reporter in at least four different fields. Nuclei are labeled with <named-entity id="named-entity-266">DAPI</named-entity>. Scale bars, 10 μm. In <b>b</b>, <b>c</b>, <b>e</b> and <b>f</b>, all values are mean ± s.e.m. Differences with control (marked with asterisk) or with serum-supplemented cells (marked with §) are significant for <i>P</i> 0.05. Full-field fluorescence images and full-length blots are shown in <sir rid="S1" refobjid="nchembio.1230-S1">Supplementary Figures 2</sir> and <sir rid="S1" refobjid="nchembio.1230-S1">21</sir>, respectively.</p>
|
https://api.sourcedata.io/file.php?figure_id=3209
|
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"ncbi_gene_id": "19401",
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"text": "RARα",
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{
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"mapping_status": "mapped",
"ncbi_gene_id": "19401",
"original_type": "gene",
"role": "intervention",
"text": "RARα",
"type": "geneprod",
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},
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q9CQV6///Q91VR7",
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"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
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"ncbi_gene_id": null,
"original_type": "protein",
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"text": "LC3",
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{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q9CQV6///Q91VR7",
"ext_tax_ids": "10090///10090",
"ext_tax_names": "Mus musculus///Mus musculus",
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{
"ext_dbs": "Uniprot///Uniprot",
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},
{
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"ext_dbs": "Uniprot///Uniprot",
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"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
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"ncbi_gene_id": null,
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},
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q9CQV6///Q91VR7",
"ext_tax_ids": "10090///10090",
"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
"mapping_status": "unmapped",
"ncbi_gene_id": null,
"original_type": "protein",
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"text": "LC3",
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{
"ext_dbs": "NCBI gene",
"ext_ids": "19401",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "19401",
"original_type": "gene",
"role": "intervention",
"text": "RARα",
"type": "geneprod",
"uniprot_ids": [
"P11416",
"Q3U3R3",
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] |
|
27110680
|
10.1038/nchembio.1230
|
Chemical modulation of chaperone-mediated autophagy by retinoic acid derivatives
|
2013
|
figf3
|
<p>(<b>a</b>) Rates of degradation of long-lived proteins in mouse fibroblasts untreated (None) or treated with 40 μM <named-entity id="named-entity-268">ATRA</named-entity> and maintained in the presence or absence of serum. Values are expressed as a percentage of proteolysis; <i>n</i> = 3. (<b>b</b>) Percentage of lysosomal degradation calculated after treatment with inhibitors of lysosomal proteolysis for 12 h; <i>n</i> = 3. (<b>c</b>) Immunoblot for <named-entity id="named-entity-269">LC3-II</named-entity> of the same cells as in <b>a</b> and <b>b</b> maintained in the presence or absence of serum and protease inhibitors (PI). Top, representative immunoblot. Actin is shown as loading control. Bottom, ratio of amount of <named-entity id="named-entity-270">LC3-II</named-entity> in cells treated with PI to that in untreated cells. Values are expressed as fold change in value relative to untreated cells; <i>n</i> = 4. 'I' and 'II' designate different forms of LC3, as described in the text. (<b>d</b>,<b>e</b>) Autophagic flux in untreated and <named-entity id="named-entity-271">ATRA</named-entity>-treated cells expressing mCherry-GFP–<named-entity id="named-entity-272">LC3-II</named-entity> and maintained in the presence or absence of serum. Shown in <b>d</b> are representative merged-channel images. Arrows indicate autolysosomes (red). Scale bar, 2 μm. Shown in <b>e</b> are the number of autophagosomes (left) and percentage of autolysosomes (right) after quantification of >50 cells. AV, autophagic vacuoles. (<b>f</b>) Mouse fibroblasts expressing the KFERQ-mCherry1 photoactivatable reporter with or without <named-entity id="named-entity-273">ATRA</named-entity> and after photoactivation maintained in the presence or absence of serum. Left, representative images. Nuclei are labeled with <named-entity id="named-entity-274">DAPI</named-entity>. Scale bars, 10 μm. Graph shows quantification of the number of puncta per cell in >50 cells. All values in <b>a</b>–<b>c</b>, <b>e</b> and <b>f</b> are mean ± s.e.m., and differences with untreated (marked with asterisk) or with serum-supplemented cells (marked with §) are significant for <i>P</i> 0.01. Full-field fluorescence images and full-length blots are shown in <sir rid="S1" refobjid="nchembio.1230-S1">Supplementary Figures 3</sir> and <sir rid="S1" refobjid="nchembio.1230-S1">21</sir>, respectively.</p>
|
https://api.sourcedata.io/file.php?figure_id=3210
|
[
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"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
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"ncbi_gene_id": null,
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"text": "LC3",
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"uniprot_ids": []
},
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q9CQV6///Q91VR7",
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"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
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"text": "LC3",
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},
{
"ext_dbs": "Uniprot///Uniprot",
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"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
"mapping_status": "unmapped",
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"original_type": "protein",
"role": "assayed",
"text": "LC3",
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"uniprot_ids": []
},
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q91VR7///Q9CQV6",
"ext_tax_ids": "10090///10090",
"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
"mapping_status": "unmapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "LC3",
"type": "geneprod",
"uniprot_ids": []
},
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q91VR7///Q9CQV6",
"ext_tax_ids": "10090///10090",
"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
"mapping_status": "unmapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "LC3",
"type": "geneprod",
"uniprot_ids": []
}
] |
|
27110680
|
10.1038/nchembio.1230
|
Chemical modulation of chaperone-mediated autophagy by retinoic acid derivatives
|
2013
|
figf5
|
<p>(<b>a</b>) Mouse fibroblasts expressing the KFERQ-mCherry1 photoactivatable reporter without (None) or with 20 μM of the indicated compounds imaged 16 h after photoactivation. Insets show higher-magnification images. Nuclei are labeled with <named-entity id="named-entity-284">DAPI</named-entity>. Scale bars, 10 μm. (<b>b</b>) Quantification of the effect of increasing concentrations of GR1 on the same cells. Untreated cells and cells treated with 40 μM <named-entity id="named-entity-285">ATRA</named-entity> are also shown. Representative images are shown in <sir rid="S1" refobjid="nchembio.1230-S1">Supplementary Figure 7</sir>. Graph shows the average number of fluorescent puncta per cell, quantified in >50 cells. All values are mean ± s.e.m. (<b>c</b>) Mouse fibroblasts were cotransfected with the human <named-entity id="named-entity-286">RARα</named-entity> (<named-entity id="named-entity-287">hRARα</named-entity>) receptor, a relevant reporter luciferase plasmid and the non–retinoid-regulated <i>Renilla</i> reporter to control for transfection. Values show relative luciferase units (RLU) detected in cells subjected to the indicated concentrations of <named-entity id="named-entity-288">ATRA</named-entity> or the three retinoid derivatives for 12 h. (<b>d</b>) Cells transfected as in <b>c</b> were treated with 100 nM of <named-entity id="named-entity-289">ATRA</named-entity> alone or in the presence of the indicated concentrations of the three retinoid derivatives or the antagonist <named-entity id="named-entity-290">BMS614</named-entity> for 12 h. Values are shown as RLU. (<b>e</b>) Mouse fibroblasts were cotransfected with the human <named-entity id="named-entity-291">RXR receptor</named-entity>, a relevant reporter luciferase plasmid and the non–retinoid-regulated <i>Renilla</i> reporter to control for transfection. Values show relative luciferase units detected in cells subjected to the indicated concentrations of <named-entity id="named-entity-292">ATRA</named-entity> or the three retinoid derivatives for 12 h. (<b>f</b>) Cells transfected as in <b>e</b> were treated with 10 μM of <named-entity id="named-entity-293">ATRA</named-entity> alone or in the presence of the indicated concentrations of the three retinoid derivatives or the antagonist <named-entity id="named-entity-294">BMS614</named-entity> for 12 h. Values are shown as RLU. Values in <b>c</b>–<b>f</b> are mean ± s.e.m.; <i>n</i> = 4–6. (<b>g</b>) Left, immunoblot for <named-entity id="named-entity-295">LC3-II</named-entity> in cells treated with 20 μM of the retinoid derivatives and protease inhibitors (PIs), as labeled. Actin is shown as loading control, and full-length blots are shown in <sir rid="S1" refobjid="nchembio.1230-S1">Supplementary Figure 21</sir>. The amount of <named-entity id="named-entity-296">LC3-II</named-entity> in untreated cells, expressed as fold change relative to control (middle), and the increase in <named-entity id="named-entity-297">LC3-II</named-entity> after PI treatment (<named-entity id="named-entity-298">LC3-II</named-entity> flux, right), expressed as fold change, were calculated from the densitometric quantification of immunoblots. Values are mean ± s.e.m.; <i>n</i> = 3.</p>
|
https://api.sourcedata.io/file.php?figure_id=3212
|
[
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"text": "RARα",
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{
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},
{
"ext_dbs": "Uniprot///Uniprot///Uniprot",
"ext_ids": "P19793///P48443///P28702",
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"ncbi_gene_id": null,
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"text": "RXR",
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},
{
"ext_dbs": "Uniprot///Uniprot///Uniprot",
"ext_ids": "P19793///P28702///P48443",
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}
] |
|
27110680
|
10.1038/nchembio.1230
|
Chemical modulation of chaperone-mediated autophagy by retinoic acid derivatives
|
2013
|
figf6
|
<p>(<b>a</b>) Rates of degradation of long-lived proteins in control mouse fibroblasts or <named-entity id="named-entity-299">RARα</named-entity>(−) or <named-entity id="named-entity-300">LAMP-2A</named-entity>(−) (<named-entity id="named-entity-301">L-2A</named-entity>(−)) cells left untreated (None) or treated with 20 μM of the indicated compounds. Values are expressed as the fold change in proteolytic rate relative to the rate in untreated cells for each group; <i>n</i> = 3. (<b>b</b>–<b>d</b>) Control mouse fibroblasts or <named-entity id="named-entity-302">RARα</named-entity>(−), <named-entity id="named-entity-303">LAMP-2A</named-entity>(−) or <named-entity id="named-entity-304">LAMP-2B</named-entity>(−) (<named-entity id="named-entity-305">L-2B</named-entity>(−)) cells were transfected with the KFERQ-mCherry1 photoactivatable reporter with or without the indicated compounds (20 μM). In <b>b</b> are shown the representative fields and high-magnification images (insets) for GR1. Nuclei are labeled with <named-entity id="named-entity-306">DAPI</named-entity>. Representative fields for GR2 and AR7 are shown in <sir rid="S1" refobjid="nchembio.1230-S1">Supplementary Figure 13</sir>. Scale bars, 10 μm. (<b>c</b>,<b>d</b>) Average number of fluorescent puncta per cell quantified in >50 cells in at least four different fields in control cells and <named-entity id="named-entity-307">RAR</named-entity>(−) (<b>c</b>) or <named-entity id="named-entity-308">LAMP-2B</named-entity>(−) (<b>d</b>) cells. No puncta were detected in <named-entity id="named-entity-309">LAMP-2A</named-entity>(−) cells. (<b>e</b>) Mouse fibroblasts transfected with the KFERQ-mCherry1 photoactivatable reporter with the indicated concentrations of AR7 or GR1 or with both compounds to reach the same final concentration, as indicated. Top, representative fields and high-magnification images (insets). Nuclei are labeled with <named-entity id="named-entity-310">DAPI</named-entity>. Bottom, quantification of the number of fluorescent puncta per field in each condition. Scale bars, 10 μm. In <b>a</b>, <b>c</b>, <b>d</b> and <b>e</b>, values are mean ± s.e.m.; <i>n</i> > 50 cells. Differences with untreated samples (marked with asterisk) or between single and combined treatments (§) are significant for <i>P</i> 0.01.</p>
|
https://api.sourcedata.io/file.php?figure_id=3213
|
[
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"text": "LAMP-2A",
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"P17047",
"Q8C5K0"
]
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"ext_tax_names": "Mus musculus",
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"Q3U3R3",
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]
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{
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]
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"P17047",
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]
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"ext_tax_names": "Mus musculus",
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"ncbi_gene_id": "16784",
"original_type": "gene",
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},
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"P17047",
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{
"ext_dbs": "NCBI gene",
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"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "19401",
"original_type": "gene",
"role": "intervention",
"text": "RARα",
"type": "geneprod",
"uniprot_ids": [
"P11416",
"Q3U3R3",
"Q3U5E7"
]
}
] |
|
27110680
|
10.1038/nchembio.1230
|
Chemical modulation of chaperone-mediated autophagy by retinoic acid derivatives
|
2013
|
figf7
|
<p>(<b>a</b>) Immunoblot for the indicated proteins in homogenates (Hom) and lysosomes (Lys) isolated from cells untreated (None) or treated for 12 h with 20 μM of the indicated compounds. (<b>b</b>) mRNA levels of <named-entity id="named-entity-311">LAMP-2A</named-entity> in control mouse fibroblasts (left) or <named-entity id="named-entity-312">RARα</named-entity>(−) cells (right) treated with AR7 or <named-entity id="named-entity-313">paraquat</named-entity> (<named-entity id="named-entity-314">PQ</named-entity>) as in <b>a</b>; <i>n</i> = 4–5. Values are presented as fold change in mRNA relative to untreated cells. (<b>c</b>) Cellular viability of control (left) or <named-entity id="named-entity-315">LAMP-2A</named-entity>(−) (right) fibroblasts exposed to 2 mM or 0.5 mM <named-entity id="named-entity-316">paraquat</named-entity>, respectively, and treated with the indicated compounds for 12 h before or after the <named-entity id="named-entity-317">paraquat</named-entity> treatment; <i>n</i> = 3. (<b>d</b>) Viability of mouse fibroblasts transfected with the indicated concentrations of a plasmid encoding <named-entity id="named-entity-318">α-synuclein</named-entity> and left untreated (None) or treated with 1 mM <named-entity id="named-entity-319">paraquat</named-entity> alone or in the presence of 20 μM AR7; <i>n</i> = 3. In <b>b</b>–<b>d</b>, all values are mean ± s.e.m. Differences with cells that are untreated (marked with asterisk) or those treated only with <named-entity id="named-entity-320">paraquat</named-entity> (§) were significant for <i>P</i> 0.001. Full-length blots are shown in <sir rid="S1" refobjid="nchembio.1230-S1">Supplementary Figure 21</sir>. (<b>e</b>) Immunoblot for <named-entity id="named-entity-321">α-synuclein</named-entity> (<named-entity id="named-entity-322">α-syn</named-entity>) in the same cells as in <b>d</b>. Top, higher-exposure blot to highlight oligomeric (oligo) species. Asterisk denotes nonspecific band. M, monomer; MW, molecular weight.</p>
|
https://api.sourcedata.io/file.php?figure_id=3214
|
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"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "16784",
"original_type": "gene",
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"text": "LAMP-2A",
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"P17047",
"Q8C5K0"
]
},
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"P11416",
"Q3U3R3",
"Q3U5E7"
]
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{
"ext_dbs": "NCBI gene",
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"ext_tax_names": "Mus musculus",
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"mapping_status": "mapped",
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"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "20617",
"original_type": "gene",
"role": "intervention",
"text": "α-synuclein",
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"ext_tax_names": "Mus musculus",
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"mapping_status": "mapped",
"ncbi_gene_id": "20617",
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"role": "intervention",
"text": "α-synuclein",
"type": "geneprod",
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"O55042"
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}
] |
|
10.15252/embj.2019101996
|
The N-end rule ubiquitin ligase UBR2 mediates NLRP1B inflammasome activation by anthrax lethal toxin
|
2019
|
Figure 1
|
<sd-panel> <p><strong>Figure 1. The N-end rule pathway controls LT-induced NLRP1B inflammasome activation.</strong></p> <p>A, B Effects of basic and bulky hydrophobic amino acids on LT-induced pyroptosis and caspase-1 activation in RAW<sup>RA</sup> cells. Cells were pre-treated with 10 mM indicated amino acids prior to LT stimulation. Cell viability was measured by using the ATP assay and data shown are mean values ± SD from three replicates (A). Mock, DMEM medium. Cell supernatants were subjected to anti-caspase-1 immunoblotting and the lysates were blotted with the anti-tubulin antibody (B). p10, mature caspase-1; p45, caspase-1 precursor.</p> <p>C, D Effects of basic and bulky hydrophobic amino acids on LT-induced RFP-ASC specks formation in RAW<sup>RA</sup> cells. Fluorescence images were taken on a confocal microscopy (C). The percentages of cells showing RFP-ASC specks (mean values ± SD from three replicates) are in (D).</p> <p>E Effects of basic and bulky hydrophobic amino acids on LT-induced caspase-1 activation in <em>Tlr4<sup>-/-</sup></em> iBMDMs. Cells were treated as that in (A). Cell supernatants were subjected to anti-caspase-1 immunoblotting and the lysates were blotted with the anti-tubulin or anti-MEK3 antibody.</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=30609
|
[
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"ext_ids": "P15917",
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"ext_tax_names": "Bacillus anthracis",
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"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "LT",
"type": "geneprod",
"uniprot_ids": [
"P15917"
]
},
{
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"ext_tax_names": "Bacillus anthracis",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "LT",
"type": "geneprod",
"uniprot_ids": [
"P15917"
]
},
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"ext_dbs": "Uniprot",
"ext_ids": "P29452",
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"ext_tax_names": "Mus musculus",
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"mapping_status": "mapped",
"ncbi_gene_id": null,
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"text": "caspase-1",
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"P29452"
]
},
{
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"text": "caspase-1",
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"P29452"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P29452",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "caspase-1",
"type": "geneprod",
"uniprot_ids": [
"P29452"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P15917",
"ext_tax_ids": "1392",
"ext_tax_names": "Bacillus anthracis",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "LT",
"type": "geneprod",
"uniprot_ids": [
"P15917"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q9EPB4",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "ASC",
"type": "geneprod",
"uniprot_ids": [
"Q9EPB4"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P29452",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "caspase-1",
"type": "geneprod",
"uniprot_ids": [
"P29452"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P29452",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "caspase-1",
"type": "geneprod",
"uniprot_ids": [
"P29452"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P15917",
"ext_tax_ids": "1392",
"ext_tax_names": "Bacillus anthracis",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "LT",
"type": "geneprod",
"uniprot_ids": [
"P15917"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O09110",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "MEK3",
"type": "geneprod",
"uniprot_ids": [
"O09110"
]
}
] |
||
10.15252/embj.2019101996
|
The N-end rule ubiquitin ligase UBR2 mediates NLRP1B inflammasome activation by anthrax lethal toxin
|
2019
|
Figure 2
|
<sd-panel> <p><strong>Figure 2. The ubiquitin ligase UBR2 is required for LT-induced NLRP1B activation.</strong></p> <p>A RFP-ASC specks formation assay of the effect of <em>Ubr2</em> knockdown on LT-induced inflammasome activation. RAW<sup>RA</sup> cells were transfected with a control siRNA or <em>Ubr2</em>-targeting siRNA mixtures for 60 h followed by LT treatment. The numbers in the merged panel are the percentages of the cells showing RFP-ASC specks.</p> <p>B Effect of <em>Ubr2</em> stable knockdown on caspase-1 activation in RAW<sup>RA</sup> cells. Cells stably expressing a control or <em>Ubr2</em>-targeting shRNA were incubated with WT LT (+) or its E687C mutant (-) for 3 h.</p> <p>C Effects of <em>Ubr2</em> knockdown (siRNA #05) on LT-induced NLRP1B inflammasome activation reconstituted in 293T cells. Cells were treated with WT LT (+) or its E687C mutant (-).</p> <p>D, E Effect of <em>Ubr2</em> knockout on NLRP1B or NAIP2-NLRC4 inflammasome activation. WT and <em>Ubr2</em><sup>-/-</sup> iBMDMs were treated with LT or BsaK for 3 h. Cell viability was measured by using the ATP assay and data shown are mean values ± SD from three replicates (D).</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=30610
|
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"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "224826",
"original_type": "gene",
"role": "intervention",
"text": "Ubr2",
"type": "geneprod",
"uniprot_ids": [
"Q6WKZ8",
"Q3UPU3"
]
},
{
"ext_dbs": "NCBI gene",
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"mapping_status": "mapped",
"ncbi_gene_id": "224826",
"original_type": "gene",
"role": "intervention",
"text": "Ubr2",
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"uniprot_ids": [
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"Q3UPU3"
]
},
{
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"ext_tax_names": "Bacillus anthracis",
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{
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},
{
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"P15917"
]
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"ext_tax_names": "Homo sapiens",
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"mapping_status": "mapped",
"ncbi_gene_id": "23304",
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"text": "Ubr2",
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"Q8IWV8",
"B3KXG6"
]
},
{
"ext_dbs": "Uniprot",
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"ext_tax_names": "Bacillus anthracis",
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"ext_dbs": "NCBI gene",
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"text": "Ubr2",
"type": "geneprod",
"uniprot_ids": [
"Q6WKZ8",
"Q3UPU3"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "A0A069BGK0",
"ext_tax_ids": "28450",
"ext_tax_names": "Burkholderia pseudomallei",
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"text": "BsaK",
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"uniprot_ids": [
"A0A069BGK0"
]
},
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] |
||
10.15252/embj.2019101996
|
The N-end rule ubiquitin ligase UBR2 mediates NLRP1B inflammasome activation by anthrax lethal toxin
|
2019
|
Figure 3
|
<sd-panel> <p><strong>Figure 3. UBE2O functions together with UBR2 to mediate NLRP1B inflammasome activation.</strong></p> <p>A Immunoblotting of caspase-1 activation in Ubrs-knockdown BMDMs derived from the 129/Sv mice. Cells were transfected with the indicated siRNA for 60 h, and then treated with WT LT (+) or its E687C mutant (-) for 3 h.</p> <p>B-D Immunoblotting of caspase-1 activation in E2s-knockdown BMDMs derived from the 129/Sv mice. Additional <em>Ube2o</em> siRNAs were used to confirm the inhibitory effect of <em>Ube2o</em> knockdown on LT-induced caspase-1 activation (C). <em>Ube2o</em> knockdown efficiency was measured by qPCR and are shown as mean values ± SD from three replicates.</p> <p>E RFP-ASC specks formation assay of the effect of <em>Ube2o</em> knockdown (siRNA <em>Ube2o</em>-01) on LT-induced inflammasome activation in RAW<sup>RA</sup> cells. The percentages of cells showing RFP-ASC specks (mean values ± SD from three replicates) are in (D).</p> <p>F Co-immunoprecipitation interaction of UBE2O and UBR2. Myc-UBE2O and Flag-UBR2 were co-expressed in 293T cells and cell lysates were subjected to anti-Flag immunoprecipitation followed by immunoblotting analyses as shown.</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=30611
|
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"ncbi_gene_id": null,
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"text": "caspase-1",
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]
},
{
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"P15917"
]
},
{
"ext_dbs": "NCBI gene",
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]
},
{
"ext_dbs": "NCBI gene",
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]
},
{
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]
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"ext_ids": "Q9EPB4",
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"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
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"text": "ASC",
"type": "geneprod",
"uniprot_ids": [
"Q9EPB4"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "224826",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "224826",
"original_type": "gene",
"role": "intervention",
"text": "UBR2",
"type": "geneprod",
"uniprot_ids": [
"Q6WKZ8",
"Q3UPU3"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q6WKZ8",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "UBR2",
"type": "geneprod",
"uniprot_ids": [
"Q6WKZ8"
]
}
] |
||
10.15252/embj.2019101996
|
The N-end rule ubiquitin ligase UBR2 mediates NLRP1B inflammasome activation by anthrax lethal toxin
|
2019
|
Figure 4
|
<sd-panel> <p><strong>Figure 4. LT cleavage of NLRP1B triggers N-end rule-mediated degradation of NLRP1B for inflammasome activation.</strong></p> <p>A Domain structure of NLRP1B. The LT cleavage site and the FIIND auto-cleavage site are marked.</p> <p>B Effect of LT treatment on NLRP1B protein level. 293T cells were transfected with Flag-NLRP1B expression plasmid for 24 h, and cells were then treated with WT LT (+) or its E687C mutant (-) for 4 h. Shown is the anti-Flag immunoblot of the total cell lysates.</p> <p>C RFP-ASC specks formation in 293T cells expressing an indicated NLRP1B variant. Cells were treated with LT for 3 h. Scale bar, 20 μm. 1-983 and 984-C (residues 984 to the C terminus) are the FIIND-mediated autocleaved fragments of NLRP1B.</p> <p>D, E Effect of MG132 and Leu on LT-induced NLRP1B degradation. 293T cells expressing NLRP1B (1-983)-HA were treated with LT and MG132 for 8 h. 10 mM Leu was added to cells 30 min prior to LT treatment. Shown are the anti-HA and anti-tubulin immunoblots of the total cell lysates.</p> <p>F LT-induced ubiquitination of NLRP1B. 293T cells expressing NLRP1B (1-983)-HA were treated with LT and/or MG132 for 6 h. Cells were then harvested for anti-HA immunoprecipitation and subjected to immunoblotting analyses using indicated antibodies.</p> <p>Expanded View Figure Legends</p> </sd-panel>
|
https://api.sourcedata.io/file.php?figure_id=30612
|
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"text": "LT",
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]
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]
},
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"ext_ids": "Q2LKW6",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
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"uniprot_ids": [
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]
}
] |
||
27520969
|
10.15252/emmm.201606413
|
IFNλ is a Potent Anti-Influenza Therapeutic without the Inflammatory Side Effects of IFNα Treatment
|
2016
|
Figure 1
|
<p><strong>Fig</strong><strong>ure</strong><strong> 1. Therapeutic administration of IFN</strong><strong>α and IFNλ</strong><strong> differentially influences the outcome of IAV induced disease. </strong></p>
<p><strong>A</strong>. Relative antiviral activity of IFNα (circles) or IFNλ (triangles). AEC cultures were stimulated for 4hrs with stated IFN at specified concentrations (ng/ml) and induction of indicated ISGs was assessed by qPCR (data shown is representative of four independent experiments, n=3-4).</p>
<p><strong>B, C</strong><strong>.</strong> Mice were pretreated with equivalent doses of IFNα (1.45μg/50μl) or IFNλ (2.6μg/50μl) or Veh Ctrl (squares, 50μl PBS) 24hrs prior to infection with PR8; weight loss and survival was assessed throughout infection (B), and viral load (C) assessed at 4dpi (data shown is representative of two independent experiments, n=8-10 (B), n=3 (C)).</p>
<p><strong>D, E</strong><strong>.</strong> Mice were infected with PR8 and treated with equivalent doses of IFNα or IFNλ or Veh Ctrl at days 2, 4 and 5 post infection; survival and weight loss was monitored (D, data pooled from 4 independent experiments, n=12-29) and viral load assessed at 4dpi (E) (data representative of two independent experiments, n=3-5).</p>
<p>Data information: Significance assessed by Log-rank (Mantel-Cox) test (survival), 2-way ANOVA (weight loss) and Unpaired t tests (viral load). * indicates IFNα:Veh Ctrl, <sup>+</sup> indicates IFNλ:Veh Ctrl, and ° indicates IFNα:IFNλ. * P=0.0236, <sup>+</sup> P=0.0236 *** P<0.0001, <sup>+++</sup> P<0.001 (B); * P= 0.012, <sup>+</sup> P=0.012 (C); * P=0.0443, <sup>+</sup> P=0.035, °° P=0.0015 (D); ** P=0.0081, <sup>++</sup> P=0.0066 (E). Symbols on the right of graphs indicate significance of whole curve. Graphs show mean ± SEM.</p>
|
https://api.sourcedata.io/file.php?figure_id=9576
|
[
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"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
"mapping_status": "unmapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNλ",
"type": "geneprod",
"uniprot_ids": []
},
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q8CGK6///Q4VK74",
"ext_tax_ids": "10090///10090",
"ext_tax_names": "Mus musculus///Mus musculus",
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},
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q8CGK6///Q4VK74",
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"ext_tax_names": "Mus musculus///Mus musculus",
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"role": "intervention",
"text": "IFNλ",
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},
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q8CGK6///Q4VK74",
"ext_tax_ids": "10090///10090",
"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
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"role": "intervention",
"text": "IFNλ",
"type": "geneprod",
"uniprot_ids": []
}
] |
|
27520969
|
10.15252/emmm.201606413
|
IFNλ is a Potent Anti-Influenza Therapeutic without the Inflammatory Side Effects of IFNα Treatment
|
2016
|
Figure 2
|
<p><strong>Fig</strong><strong>ure</strong><strong> 2. IFNα treatment correlates with increased inflammation during IAV infection. </strong></p>
<p><strong>A, B</strong><strong>.</strong> Mice were infected with PR8 and treated with IFNα (circles, 1.45μg/50μl), IFNλ (triangles, 2.6μg/50μl) or Veh Ctrl (squares) as previously stated. Concentrations of stated proinflammatory cytokines in BAL fluid was measured by multiplex cytokine assay (A) and flow cytometric quantification of pDCs and Inflammatory Monocytes in the lung was performed (B) (data shown is representative of two independent experiments, n=2-6).</p>
<p><strong>C, D</strong><strong>.</strong> Lung sections from control and infected mice treated as indicated, were stained by TUNEL for apoptotic cells at 6dpi. Quantification of TUNEL+ cells in whole lung slides by Icy Spot Detector (ICY-R3M2Y2) (C) (data shown is pooled from three independent experiments, n=3-8). Red arrowheads indicate TUNEL signal (D). Scale bar, 200 μM (data shown is representative of two independent experiments, n=3-4).</p>
<p>Data information: Significance assessed by 2-way ANOVA with Bonferroni post tests (where * denotes IFNα:Veh Ctrl, <sup>+</sup> indicates IFNλ:Veh Ctrl , and ° indicates IFNα:IFNλ). Symbols on the right of graphs indicate statistical significance of the whole curve. IL-6 whole curve: ** 0.0041, ° P=0.0144. IL-6 5 d.p.i.: ** P=0.001884, °° P=0.001645. IP-10 5 d.p.i.: ** P=0.004897, °° P=0.005354. MCP-1 5.d.p.i.: ** P=0.007473, °° P= 0.002003. Eotaxin whole curve: * 0.0235, ° P=0.0386. Eotaxin 5 d.p.i. *** P=0.000149, °° P=0.001975. Mip-1α 5 d.p.i.: °° P= 0.002921 (A). Plasmacytoid Dendritic Cells: * P=0.0211, °° P= 0.006965. Inflammatory monocytes ** P=0.007842, °° P=0.000895 (B). ** P=0.0011, <sup>++</sup> P=0.0051, °°° P=0.0005 (C). Graphs show mean ± SEM.</p>
|
https://api.sourcedata.io/file.php?figure_id=9577
|
[
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q8CGK6///Q4VK74",
"ext_tax_ids": "10090///10090",
"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
"mapping_status": "unmapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNλ",
"type": "geneprod",
"uniprot_ids": []
},
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q8CGK6///Q4VK74",
"ext_tax_ids": "10090///10090",
"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
"mapping_status": "unmapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNλ",
"type": "geneprod",
"uniprot_ids": []
}
] |
|
27520969
|
10.15252/emmm.201606413
|
IFNλ is a Potent Anti-Influenza Therapeutic without the Inflammatory Side Effects of IFNα Treatment
|
2016
|
Figure 3
|
<p><strong>Fig</strong><strong>ure</strong><strong> 3. IFNα, but not IFNλ treatment induces </strong><strong>pulmonary </strong><strong>cytokine secretion through activation of immune cells. </strong></p>
<p><strong>A, B</strong><strong>.</strong> IL-6, IP-10 and MCP-1 concentrations were measured by multiplex cytokine assay in AEC culture supernatants (A) and Macrophage, pDC and cDC culture supernatants (B) at 24hrs post stimulation with IFNα4 (0.725ng/ml) or IFNλ2 (1.3ng/ml) or LPS (AEC only) (data shown is representative of two independent experiments, n=3-6).</p>
<p><strong>C</strong>. BAL samples taken from mice treated with IFNα, IFNλ or Veh Ctrl at specified time points (data shown is representative of two independent experiments, n=5-6).</p>
<p>Data information: Significance assessed by Unpaired t tests where * denotes IFNα:Veh Ctrl and ° indicates IFNα:IFNλ. IFNλ:Veh Ctrl was not significant. IL6 pDC: *** P=0.0004, °°° P= 0.0005, IL6 cDC: * P=0.0102, ° P=0.0151. IP-10 macrophage: ** P=0.0033, °° P=0.0033, pDC: **** P<0.0001, °°°° P<0.0001, cDC: ** P= 0.0013, °° P=0.0013, MCP-1 macrophage: **** P<0.0001, °°° P=0.003 (B). IL-6 10hrs: * P=0.0112, ° P=0.0262, 18hrs: * P=0.0314, ° P=0.373. IP-10 10hrs: * P= 0.0261, ° P=0.0472. MCP-1 10hrs: ** P=0.0081, ° P=0.0206, 18hrs: ** P=0.0089, ° P=0.01 (C).</p>
|
https://api.sourcedata.io/file.php?figure_id=9578
|
[
{
"ext_dbs": "Uniprot",
"ext_ids": "P10148",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "MCP-1",
"type": "geneprod",
"uniprot_ids": [
"P10148"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P17515",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "IP-10",
"type": "geneprod",
"uniprot_ids": [
"P17515"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P07351",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNα4",
"type": "geneprod",
"uniprot_ids": [
"P07351"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q4VK74",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNλ2",
"type": "geneprod",
"uniprot_ids": [
"Q4VK74"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P08505",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "IL-6",
"type": "geneprod",
"uniprot_ids": [
"P08505"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P10148",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "MCP-1",
"type": "geneprod",
"uniprot_ids": [
"P10148"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P17515",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "IP-10",
"type": "geneprod",
"uniprot_ids": [
"P17515"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P07351",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNα4",
"type": "geneprod",
"uniprot_ids": [
"P07351"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q4VK74",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNλ2",
"type": "geneprod",
"uniprot_ids": [
"Q4VK74"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P08505",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "IL-6",
"type": "geneprod",
"uniprot_ids": [
"P08505"
]
},
{
"ext_dbs": "Uniprot///Uniprot",
"ext_ids": "Q8CGK6///Q4VK74",
"ext_tax_ids": "10090///10090",
"ext_tax_names": "Mus musculus///Mus musculus",
"mapping_source": "unknown",
"mapping_status": "unmapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNλ",
"type": "geneprod",
"uniprot_ids": []
}
] |
|
27520969
|
10.15252/emmm.201606413
|
IFNλ is a Potent Anti-Influenza Therapeutic without the Inflammatory Side Effects of IFNα Treatment
|
2016
|
Figure 4
|
<p><strong>Fig</strong><strong>ure</strong><strong> 4. Pathogenicity-related gene clusters are specifically induced by IFNα, not by IFNλ treatment. </strong></p>
<p>Mice were treated with IFNα (1.45μg/50μl), IFNλ (2.6μg/50μl) or Veh Ctrl (50μl PBS), and whole lungs were taken at 18hrs post treatment for global analysis by Illumina.SingleColor.Mouse WG-6_V2_0_R0_1127 microarrays. Samples (n=5) were normalized to the median of the vehicle control group and filtered for a fold change of 1.5, yielding 553 genes differently regulated between treatments (One way ANOVA, P<0.01, Benjamini-Hochberg multiple test correction), of which 429 genes are upregulated. K means clustering revealed six gene clusters, one of which encompassed genes primarily induced by IFNα4 (A), while the remaining clusters contained genes upregulated by both IFNα4 and IFNλ2 (B). The two clusters of genes were analysed by Ingenuity Pathway Analysis (IPA) (C,D).</p>
|
https://api.sourcedata.io/file.php?figure_id=9579
|
[
{
"ext_dbs": "Uniprot",
"ext_ids": "P07351",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNα4",
"type": "geneprod",
"uniprot_ids": [
"P07351"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P07351",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNα4",
"type": "geneprod",
"uniprot_ids": [
"P07351"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q4VK74",
"ext_tax_ids": "10090",
"ext_tax_names": "Mus musculus",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNλ2",
"type": "geneprod",
"uniprot_ids": [
"Q4VK74"
]
}
] |
|
27520969
|
10.15252/emmm.201606413
|
IFNλ is a Potent Anti-Influenza Therapeutic without the Inflammatory Side Effects of IFNα Treatment
|
2016
|
Figure 5
|
<p><strong>Fig</strong><strong>ure</strong><strong> 5. IFNα, but not IFNλ treatment induces cytokine secretion from human immune cells. </strong></p>
<p><strong>A</strong>. Human AEC cultures were stimulated for 4hrs with IFNα (circles) or IFNλ (triangles) at specified concentrations then assessed for stated ISG induction by qPCR (data is representative of 2 independent experiments, n=3).</p>
<p><strong>B</strong><strong>, C</strong><strong>.</strong> ISG induction in human PBMCs was assessed at 4 and 24hrs post IFNα (21 U/ml) or IFNλ (1.2 ng/ml) stimulation (B). PBMC proinflammatory cytokine secretion was measured by multiplex cytokine assay at 4 and 24hrs post stimulation with IFNα or IFNλ (C) (data shown is pooled from 6 independent donors).</p>
<p>Data information: Significance tested by 2-way ANOVA where * denotes IFNα:Veh Ctrl and ° indicates IFNα:IFNλ. IFNλ:Veh Ctrl was not significant. IRF7: ** P=0.0037, Rsad2 4hrs: ** P=0.0038, °° P=0.037, 24hrs: * P=0.0234. OAS1 4hrs: * P=0.0328, ° P=0.0358, 24hrs: **** P<0.0001 (B). IL-6: * P=0.0124, ** P=0.0021. MCP-1 4hrs: **** P<0.0001, °°° P=0.001, 24hrs: ** P=0.0046, °° P= 0.005. IP-10 4hrs: ** P=0.0033, °° P= 0.0024, 24hrs: ** P=0.0011, °°° P=0.001 (C).</p>
|
https://api.sourcedata.io/file.php?figure_id=9580
|
[
{
"ext_dbs": "Uniprot///Uniprot///Uniprot///Uniprot",
"ext_ids": "Q8IZI9///K9M1U5///Q8IZJ0///Q8IU54",
"ext_tax_ids": "9606///9606///9606///9606",
"ext_tax_names": "Homo sapiens///Homo sapiens///Homo sapiens///Homo sapiens",
"mapping_source": "unknown",
"mapping_status": "unmapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNλ",
"type": "geneprod",
"uniprot_ids": []
},
{
"ext_dbs": "Uniprot///Uniprot///Uniprot///Uniprot",
"ext_ids": "Q8IZI9///K9M1U5///Q8IZJ0///Q8IU54",
"ext_tax_ids": "9606///9606///9606///9606",
"ext_tax_names": "Homo sapiens///Homo sapiens///Homo sapiens///Homo sapiens",
"mapping_source": "unknown",
"mapping_status": "unmapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNλ",
"type": "geneprod",
"uniprot_ids": []
},
{
"ext_dbs": "Uniprot///Uniprot///Uniprot///Uniprot",
"ext_ids": "Q8IZI9///K9M1U5///Q8IZJ0///Q8IU54",
"ext_tax_ids": "9606///9606///9606///9606",
"ext_tax_names": "Homo sapiens///Homo sapiens///Homo sapiens///Homo sapiens",
"mapping_source": "unknown",
"mapping_status": "unmapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "intervention",
"text": "IFNλ",
"type": "geneprod",
"uniprot_ids": []
}
] |
|
27596438
|
10.15252/emmm.201505971
|
ZEB1-mediated melanoma cell plasticity enhances resistance to MAPK inhibitors
|
2016
|
Figure 1
|
<p><strong>Figure 1</strong><strong>.</strong> <strong>High </strong><strong>levels of </strong><strong>ZEB1 expression </strong><strong>are </strong><strong>correlate</strong><strong>d</strong><strong> with low </strong><strong>MITF</strong><strong> level</strong><strong>s</strong><strong> and</strong> <strong>are</strong><strong> associated with inherent resistance to </strong><strong>MAPKi</strong><strong> in </strong><strong><em>BRAF</em></strong><strong><em><sup>V600</sup></em></strong><strong>-mutated </strong><strong>melanoma cell lines</strong></p>
<p>(A) Pearson correlation between <em>ZEB1</em> and <em>MITF</em> mRNA expression in 61 melanoma cell lines available through the CCLE. (B) ZEB1, ZEB2, TWIST1 and MITF expression in a panel of <em>BRAF</em><em><sup>V600</sup></em>-mutated melanoma cells assessed by Western blot. GLO and C-09.10 cells are patient-derived short-term cultures. Actin was used as a loading control. (C) Quantitative PCR analyses of <em>ZEB1</em>, <em>ZEB2</em>, <em>TWIST1 </em>and <em>MITF</em> in the same panel of cell lines. mRNA expression levels are represented relatively to C-09.10 cells, in which the levels were arbitrarily fixed at 1 (n=3, mean ± SD). The dotted line separates ZEB1<sup>high</sup> (left) and ZEB1<sup>low</sup> (right) cell lines. (D) Tukey box plot of <em>ZEB1</em>, <em>ZEB2</em>, <em>TWIST1</em>, and <em>MITF</em> mRNA expression according to the IC50 of the drug (µM) administered (BRAFi/MEKi), in melanoma cell lines from the CCLE (n=28) (Student’s t-Test). High <em>ZEB1</em>, low <em>ZEB2</em>, and low <em>MITF</em> expression levels were correlated with BRAFi (PLX4720) and MEKi (AZD6244) resistance. PLX4720 is an analog of PLX4032. (E) IC50 values of PLX4032 (µM) in the panel of <em>BRAF</em><em><sup>V600</sup></em> melanoma cells as determined by ATP assay (n=3, mean ± SD). For SKMEL24 and WM793, IC50 was >8µM.</p>
|
https://api.sourcedata.io/file.php?figure_id=9817
|
[
{
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"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "4286",
"original_type": "gene",
"role": "assayed",
"text": "MITF",
"type": "geneprod",
"uniprot_ids": [
"O75030",
"A0A087WXU1",
"A0A8I5KSZ4",
"B4DNC7"
]
},
{
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"mapping_status": "mapped",
"ncbi_gene_id": "6935",
"original_type": "gene",
"role": "assayed",
"text": "ZEB1",
"type": "geneprod",
"uniprot_ids": [
"P37275",
"B2RBI8",
"B4DGU2"
]
},
{
"ext_dbs": "Uniprot",
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"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "MITF",
"type": "geneprod",
"uniprot_ids": [
"O75030"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q15672",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "TWIST1",
"type": "geneprod",
"uniprot_ids": [
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]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P37275",
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"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
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"text": "ZEB1",
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]
},
{
"ext_dbs": "Uniprot",
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"text": "ZEB2",
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"uniprot_ids": [
"O60315"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "4286",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "4286",
"original_type": "gene",
"role": "assayed",
"text": "MITF",
"type": "geneprod",
"uniprot_ids": [
"O75030",
"A0A087WXU1",
"A0A8I5KSZ4",
"B4DNC7"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "6935",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "6935",
"original_type": "gene",
"role": "assayed",
"text": "ZEB1",
"type": "geneprod",
"uniprot_ids": [
"P37275",
"B2RBI8",
"B4DGU2"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "6935",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "6935",
"original_type": "gene",
"role": "assayed",
"text": "ZEB1",
"type": "geneprod",
"uniprot_ids": [
"P37275",
"B2RBI8",
"B4DGU2"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "6935",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "6935",
"original_type": "gene",
"role": "assayed",
"text": "ZEB1",
"type": "geneprod",
"uniprot_ids": [
"P37275",
"B2RBI8",
"B4DGU2"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "4286",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "4286",
"original_type": "gene",
"role": "assayed",
"text": "MITF",
"type": "geneprod",
"uniprot_ids": [
"O75030",
"A0A087WXU1",
"A0A8I5KSZ4",
"B4DNC7"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "4286",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "4286",
"original_type": "gene",
"role": "assayed",
"text": "MITF",
"type": "geneprod",
"uniprot_ids": [
"O75030",
"A0A087WXU1",
"A0A8I5KSZ4",
"B4DNC7"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "7291",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "7291",
"original_type": "gene",
"role": "assayed",
"text": "TWIST1",
"type": "geneprod",
"uniprot_ids": [
"Q15672"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "6935",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "6935",
"original_type": "gene",
"role": "assayed",
"text": "ZEB1",
"type": "geneprod",
"uniprot_ids": [
"P37275",
"B2RBI8",
"B4DGU2"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "6935",
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"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
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"P37275",
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"B4DGU2"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "9839",
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"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
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"original_type": "gene",
"role": "assayed",
"text": "ZEB2",
"type": "geneprod",
"uniprot_ids": [
"O60315"
]
},
{
"ext_dbs": "NCBI gene",
"ext_ids": "9839",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "9839",
"original_type": "gene",
"role": "assayed",
"text": "ZEB2",
"type": "geneprod",
"uniprot_ids": [
"O60315"
]
}
] |
|
27596438
|
10.15252/emmm.201505971
|
ZEB1-mediated melanoma cell plasticity enhances resistance to MAPK inhibitors
|
2016
|
Figure 2
|
<p><strong>Figure </strong><strong>2</strong><strong>.</strong> <strong>High ZEB1 </strong><strong>and </strong><strong>low </strong><strong>MITF</strong><strong> level</strong><strong>s</strong> <strong>are</strong><strong> associated with inherent resistance to </strong><strong>MAPKi</strong><strong> in </strong><strong><em>BRAF</em></strong><strong><em><sup>V600</sup></em></strong><strong>-</strong><strong>mutated </strong><strong>melanoma </strong><strong>tumors</strong></p>
<p>(A) Pearson correlation between <em>ZEB1</em> and <em>MITF</em> mRNA expression levels in 467 melanoma tumors from TCGA data set. (B) Representative pictures of ZEB1 and MITF immunostaining in primary melanomas. Scale bar = 40 µm. The aberrant activation of ZEB1 in melanomas is correlated with a MITF<sup>low</sup> phenotype. (C) Representative pictures of ZEB1 immunostaining in <em>BRAF</em><em><sup>V600</sup></em> tumors from patients, classified in ZEB1 high, int and low subgroups, based on the intensity of ZEB1 staining and on the percentage of cells positive for ZEB1. Scale bar = 80 µm. (D) Pie charts representing the distribution of ZEB1 alone (upper part), or ZEB1 and TWIST1 (lower part) immunohistochemical staining in tumors according to their initial response to vemurafenib +/- cobimetinib treatment. ZEB1 +/- TWIST1 levels are higher in MAPKi primary resistant melanomas (initial non-responders) compared to tumors that initially respond to treatment (n=70, Fisher’s exact test). (E) Representative pictures of ZEB1 and MITF immunostainings, before and after vemurafenib treatment, in the tumor from patient 1, exhibiting primary resistance to BRAFi. Scale bar = 80 µm. Right: Magnification of MITF<sup>high</sup> and MITF<sup>low</sup> clones in the resistant tumor under treatment. Arrows indicate endothelial and stromal cells that also show positive staining for ZEB1, besides tumor cells.</p>
|
https://api.sourcedata.io/file.php?figure_id=9818
|
[
{
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"mapping_source": "ncbi_gene",
"mapping_status": "mapped",
"ncbi_gene_id": "4286",
"original_type": "gene",
"role": "assayed",
"text": "MITF",
"type": "geneprod",
"uniprot_ids": [
"O75030",
"A0A087WXU1",
"A0A8I5KSZ4",
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]
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]
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]
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{
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]
},
{
"ext_dbs": "Uniprot",
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]
},
{
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]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O75030",
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"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
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"role": "assayed",
"text": "MITF",
"type": "geneprod",
"uniprot_ids": [
"O75030"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P37275",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "ZEB1",
"type": "geneprod",
"uniprot_ids": [
"P37275"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P37275",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "ZEB1",
"type": "geneprod",
"uniprot_ids": [
"P37275"
]
}
] |
|
27596438
|
10.15252/emmm.201505971
|
ZEB1-mediated melanoma cell plasticity enhances resistance to MAPK inhibitors
|
2016
|
Figure 3
|
<p><strong>Figure </strong><strong>3</strong><strong>.</strong> <strong>ZEB1 expression is activated in </strong><strong><em>BRAF</em></strong><strong><em><sup>V600</sup></em></strong><strong>-</strong><strong>mutated </strong><strong>melanoma cell lines with acquired resistance to </strong><strong>BRAFi</strong><strong> and </strong><strong>in </strong><strong>biopsies from patients relapsing while under treatment</strong></p>
<p>(A) PLX4032 IC50 (µM) of sensitive A375 and SKMEL5 and resistant (A375-R, SKMEL5-R) cell lines, as well as of GOKA and ESP cells, two BRAFi-resistant patient-derived short-term cultures, as determined by ATP assay (n=3, mean ± SD). For ESP, IC50 was >8µM. (B) Western blot analyses of ZEB1, TWIST1 and FRA1 in A375-R and SKMEL5-R <em>versus</em> the parental naive cells, and in GOKA and ESP cells. GAPDH was used as a loading control. (C) Quantitative PCR analyses of <em>ZEB1 </em>and <em>MITF</em> in A375-R and SKMEL5-R <em>versus</em> the parental naive cells, and in GOKA and ESP cells. mRNA expression levels are represented as arbitrary units (a.u). Statistical difference relative to sensitive A375 cells is shown (n=3, mean ± SD, Student’s t-test). (D) Representative pictures of ZEB1 and MITF immunostainings in tumors from patients 2, 3 and 4, before and after vemurafenib treatment. Scale bar = 40 µm. For ZEB1 staining in patient 4, the inset shows a magnification. Arrows point at stromal cells (s). All other cells positive for ZEB1 are tumor cells.</p>
|
https://api.sourcedata.io/file.php?figure_id=9819
|
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"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "FRA1",
"type": "geneprod",
"uniprot_ids": [
"P15407"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "Q15672",
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"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "TWIST1",
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]
},
{
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]
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{
"ext_dbs": "NCBI gene",
"ext_ids": "4286",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
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"A0A087WXU1",
"A0A8I5KSZ4",
"B4DNC7"
]
},
{
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"ext_ids": "6935",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
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"ncbi_gene_id": "6935",
"original_type": "gene",
"role": "assayed",
"text": "ZEB1",
"type": "geneprod",
"uniprot_ids": [
"P37275",
"B2RBI8",
"B4DGU2"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "O75030",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "MITF",
"type": "geneprod",
"uniprot_ids": [
"O75030"
]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P37275",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
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"text": "ZEB1",
"type": "geneprod",
"uniprot_ids": [
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]
},
{
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"ext_ids": "P37275",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
"original_type": "protein",
"role": "assayed",
"text": "ZEB1",
"type": "geneprod",
"uniprot_ids": [
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]
},
{
"ext_dbs": "Uniprot",
"ext_ids": "P37275",
"ext_tax_ids": "9606",
"ext_tax_names": "Homo sapiens",
"mapping_source": "direct",
"mapping_status": "mapped",
"ncbi_gene_id": null,
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"P37275"
]
}
] |
End of preview. Expand
in Data Studio
YAML Metadata
Warning:
The task_categories "named-entity-recognition" is not in the official list: text-classification, token-classification, table-question-answering, question-answering, zero-shot-classification, translation, summarization, feature-extraction, text-generation, fill-mask, sentence-similarity, text-to-speech, text-to-audio, automatic-speech-recognition, audio-to-audio, audio-classification, audio-text-to-text, voice-activity-detection, depth-estimation, image-classification, object-detection, image-segmentation, text-to-image, image-to-text, image-to-image, image-to-video, unconditional-image-generation, video-classification, reinforcement-learning, robotics, tabular-classification, tabular-regression, tabular-to-text, table-to-text, multiple-choice, text-ranking, text-retrieval, time-series-forecasting, text-to-video, image-text-to-text, visual-question-answering, document-question-answering, zero-shot-image-classification, graph-ml, mask-generation, zero-shot-object-detection, text-to-3d, image-to-3d, image-feature-extraction, video-text-to-text, keypoint-detection, visual-document-retrieval, any-to-any, video-to-video, other
YAML Metadata
Warning:
The task_categories "named-entity-linking" is not in the official list: text-classification, token-classification, table-question-answering, question-answering, zero-shot-classification, translation, summarization, feature-extraction, text-generation, fill-mask, sentence-similarity, text-to-speech, text-to-audio, automatic-speech-recognition, audio-to-audio, audio-classification, audio-text-to-text, voice-activity-detection, depth-estimation, image-classification, object-detection, image-segmentation, text-to-image, image-to-text, image-to-image, image-to-video, unconditional-image-generation, video-classification, reinforcement-learning, robotics, tabular-classification, tabular-regression, tabular-to-text, table-to-text, multiple-choice, text-ranking, text-retrieval, time-series-forecasting, text-to-video, image-text-to-text, visual-question-answering, document-question-answering, zero-shot-image-classification, graph-ml, mask-generation, zero-shot-object-detection, text-to-3d, image-to-3d, image-feature-extraction, video-text-to-text, keypoint-detection, visual-document-retrieval, any-to-any, video-to-video, other
SODA-SPROUT: Role-Filtered Named Entity Linking Dataset
Dataset Description
This dataset is an improved version of the SODA-SPROUT NEL dataset, specifically filtered to include only the most relevant biological entities for Named Entity Linking tasks. The dataset focuses on proteins and genes with roles of 'assayed' and 'intervention', which represent the core biological entities that are actually measured or manipulated in scientific experiments.
Key Improvements
- Role Filtering: Only includes entities with roles 'assayed' (104,286 entities) and 'intervention' (80,127 entities)
- Higher Quality: Removes less relevant entities like 'reporter', 'component', 'normalizing', and 'experiment' roles
- Better NEL Performance: Focuses on the most important biological entities for linking tasks
- Reduced Noise: 16,532 figure-level records (vs. 21,000+ in unfiltered version)
Dataset Structure
Statistics
- Total Records: 16,532 figure-level records
- Training Set: 14,880 records (90%)
- Validation Set: 825 records (5%)
- Test Set: 827 records (5%)
- Total Entities: 184,413 (104,286 assayed + 80,127 intervention)
- Unique Papers: 3,101 papers
Data Format
Each record contains:
- Paper Context: PMID, DOI, title, abstract, year
- Figure Context: Figure label, caption, figure URL
- Entities: List of gene/protein entities with:
text: Entity mention texttype: "geneprod" (for NER compatibility)original_type: "gene" or "protein" (from SourceData)role: "assayed" or "intervention"ext_tax_names: Organism taxonomy nameuniprot_ids: Ground truth UniProt accessionsmapping_status: "mapped" or "unmapped"
Usage
from datasets import load_dataset
# Load the dataset
dataset = load_dataset("EMBO/soda-nel")
# Access splits
train_data = dataset["train"]
val_data = dataset["validation"]
test_data = dataset["test"]
# Example record
record = train_data[0]
print(f"Paper: {record['title']}")
print(f"Figure: {record['fig_label']}")
print(f"Caption: {record['caption']}")
print(f"Entities: {len(record['entities'])}")
Role Definitions
- Assayed: Proteins/genes that were actually measured, quantified, or tested in the experiment
- Intervention: Proteins/genes that were manipulated, targeted, or used as experimental interventions
Data Sources
- Source Dataset: EMBO/SourceData
- Original Paper: SourceData: A platform for the large-scale curation of unstructured data from the scientific literature
- UniProt Mapping: NCBI Gene IDs mapped to UniProt accessions via UniProt REST API
Citation
If you use this dataset, please cite both the original SourceData paper and this improved version:
@article{source_data_2023,
title={SourceData: A platform for the large-scale curation of unstructured data from the scientific literature},
author={...},
journal={...},
year={2023},
url={https://arxiv.org/abs/2310.20440}
}
License
This dataset is released under the MIT License. The underlying SourceData is available under its original license.
Updates
- v1.1.0: Added role filtering for 'assayed' and 'intervention' entities only
- v1.0.0: Initial release with all entity roles
- Downloads last month
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