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anti april  (Miltenyi Biotec)
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( A ) ELISA for secreted TNFSF13 in colonoid conditioned media ( n = 3 patient lines for control and VEO-IBD; n = 3 passages for variant). ( B ) Representative TNFSF13 RNAscope in colonoids from control and variant participants (scale bar: 50 μm; n = 3 patient lines for control; n = 3 passages for variant). ( C ) Costaining of TNFSF13 and FAS RNAscope probes with Ki67 antibody in colon biopsies. Arrows indicate cells accumulated outside epithelial crypts ( n = 3 patients for control; n = 3 tissue blocks for variant). ( D ) Representative images of colonoid formation assays at day 6 after seeding (scale bar: 300 μm). ( E ) Quantification of newly formed colonoids by size at day 6. Each passage included 2 technical replicates ( n = 4 patient lines for control and VEO-IBD; n = 4 passages for variant). ( F ) TNFSF13 and FAS RNAscope with E-cadherin immunostaining in WT and variant iPSC-derived colon organoids at day 7 (scale bar: 50 μm; n = 3 passages). ( G ) Representative colonoid formation in WT and variant iPSC-derived organoids at day 9 (scale bar: 400 μm; n = 3 passages, each with ≥ 2 technical replicates). ( H ) Quantification of colonoid formation rate and area at day 9. Colonoid size calculated by maximum vertical projection area. ( I and J ) Percentage of EdU + cells following IgG or TNFSF13 neutralizing antibody <t>(nTNFSF13)</t> treatment in ( I ) control tissue–derived colonoids ( n = 3 patient lines) or ( J ) WT iPSC-organoids at day 7 ( n = 3 passages). 2-way ANOVA with multiple comparisons was used for ( A and E ); 2-tailed Student’s t test for ( H – J ). P values shown on graphs unless P > 0.05.
Ntnfsf13, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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( A ) ELISA for secreted TNFSF13 in colonoid conditioned media ( n = 3 patient lines for control and VEO-IBD; n = 3 passages for variant). ( B ) Representative TNFSF13 RNAscope in colonoids from control and variant participants (scale bar: 50 μm; n = 3 patient lines for control; n = 3 passages for variant). ( C ) Costaining of TNFSF13 and FAS RNAscope probes with Ki67 antibody in colon biopsies. Arrows indicate cells accumulated outside epithelial crypts ( n = 3 patients for control; n = 3 tissue blocks for variant). ( D ) Representative images of colonoid formation assays at day 6 after seeding (scale bar: 300 μm). ( E ) Quantification of newly formed colonoids by size at day 6. Each passage included 2 technical replicates ( n = 4 patient lines for control and VEO-IBD; n = 4 passages for variant). ( F ) TNFSF13 and FAS RNAscope with E-cadherin immunostaining in WT and variant iPSC-derived colon organoids at day 7 (scale bar: 50 μm; n = 3 passages). ( G ) Representative colonoid formation in WT and variant iPSC-derived organoids at day 9 (scale bar: 400 μm; n = 3 passages, each with ≥ 2 technical replicates). ( H ) Quantification of colonoid formation rate and area at day 9. Colonoid size calculated by maximum vertical projection area. ( I and J ) Percentage of EdU + cells following IgG or TNFSF13 neutralizing antibody <t>(nTNFSF13)</t> treatment in ( I ) control tissue–derived colonoids ( n = 3 patient lines) or ( J ) WT iPSC-organoids at day 7 ( n = 3 passages). 2-way ANOVA with multiple comparisons was used for ( A and E ); 2-tailed Student’s t test for ( H – J ). P values shown on graphs unless P > 0.05.
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( A ) ELISA for secreted TNFSF13 in colonoid conditioned media ( n = 3 patient lines for control and VEO-IBD; n = 3 passages for variant). ( B ) Representative TNFSF13 RNAscope in colonoids from control and variant participants (scale bar: 50 μm; n = 3 patient lines for control; n = 3 passages for variant). ( C ) Costaining of TNFSF13 and FAS RNAscope probes with Ki67 antibody in colon biopsies. Arrows indicate cells accumulated outside epithelial crypts ( n = 3 patients for control; n = 3 tissue blocks for variant). ( D ) Representative images of colonoid formation assays at day 6 after seeding (scale bar: 300 μm). ( E ) Quantification of newly formed colonoids by size at day 6. Each passage included 2 technical replicates ( n = 4 patient lines for control and VEO-IBD; n = 4 passages for variant). ( F ) TNFSF13 and FAS RNAscope with E-cadherin immunostaining in WT and variant iPSC-derived colon organoids at day 7 (scale bar: 50 μm; n = 3 passages). ( G ) Representative colonoid formation in WT and variant iPSC-derived organoids at day 9 (scale bar: 400 μm; n = 3 passages, each with ≥ 2 technical replicates). ( H ) Quantification of colonoid formation rate and area at day 9. Colonoid size calculated by maximum vertical projection area. ( I and J ) Percentage of EdU + cells following IgG or TNFSF13 neutralizing antibody <t>(nTNFSF13)</t> treatment in ( I ) control tissue–derived colonoids ( n = 3 patient lines) or ( J ) WT iPSC-organoids at day 7 ( n = 3 passages). 2-way ANOVA with multiple comparisons was used for ( A and E ); 2-tailed Student’s t test for ( H – J ). P values shown on graphs unless P > 0.05.
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( A ) ELISA for secreted TNFSF13 in colonoid conditioned media ( n = 3 patient lines for control and VEO-IBD; n = 3 passages for variant). ( B ) Representative TNFSF13 RNAscope in colonoids from control and variant participants (scale bar: 50 μm; n = 3 patient lines for control; n = 3 passages for variant). ( C ) Costaining of TNFSF13 and FAS RNAscope probes with Ki67 antibody in colon biopsies. Arrows indicate cells accumulated outside epithelial crypts ( n = 3 patients for control; n = 3 tissue blocks for variant). ( D ) Representative images of colonoid formation assays at day 6 after seeding (scale bar: 300 μm). ( E ) Quantification of newly formed colonoids by size at day 6. Each passage included 2 technical replicates ( n = 4 patient lines for control and VEO-IBD; n = 4 passages for variant). ( F ) TNFSF13 and FAS RNAscope with E-cadherin immunostaining in WT and variant iPSC-derived colon organoids at day 7 (scale bar: 50 μm; n = 3 passages). ( G ) Representative colonoid formation in WT and variant iPSC-derived organoids at day 9 (scale bar: 400 μm; n = 3 passages, each with ≥ 2 technical replicates). ( H ) Quantification of colonoid formation rate and area at day 9. Colonoid size calculated by maximum vertical projection area. ( I and J ) Percentage of EdU + cells following IgG or TNFSF13 neutralizing antibody <t>(nTNFSF13)</t> treatment in ( I ) control tissue–derived colonoids ( n = 3 patient lines) or ( J ) WT iPSC-organoids at day 7 ( n = 3 passages). 2-way ANOVA with multiple comparisons was used for ( A and E ); 2-tailed Student’s t test for ( H – J ). P values shown on graphs unless P > 0.05.
Cell Removal Beads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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( A ) ELISA for secreted TNFSF13 in colonoid conditioned media ( n = 3 patient lines for control and VEO-IBD; n = 3 passages for variant). ( B ) Representative TNFSF13 RNAscope in colonoids from control and variant participants (scale bar: 50 μm; n = 3 patient lines for control; n = 3 passages for variant). ( C ) Costaining of TNFSF13 and FAS RNAscope probes with Ki67 antibody in colon biopsies. Arrows indicate cells accumulated outside epithelial crypts ( n = 3 patients for control; n = 3 tissue blocks for variant). ( D ) Representative images of colonoid formation assays at day 6 after seeding (scale bar: 300 μm). ( E ) Quantification of newly formed colonoids by size at day 6. Each passage included 2 technical replicates ( n = 4 patient lines for control and VEO-IBD; n = 4 passages for variant). ( F ) TNFSF13 and FAS RNAscope with E-cadherin immunostaining in WT and variant iPSC-derived colon organoids at day 7 (scale bar: 50 μm; n = 3 passages). ( G ) Representative colonoid formation in WT and variant iPSC-derived organoids at day 9 (scale bar: 400 μm; n = 3 passages, each with ≥ 2 technical replicates). ( H ) Quantification of colonoid formation rate and area at day 9. Colonoid size calculated by maximum vertical projection area. ( I and J ) Percentage of EdU + cells following IgG or TNFSF13 neutralizing antibody <t>(nTNFSF13)</t> treatment in ( I ) control tissue–derived colonoids ( n = 3 patient lines) or ( J ) WT iPSC-organoids at day 7 ( n = 3 passages). 2-way ANOVA with multiple comparisons was used for ( A and E ); 2-tailed Student’s t test for ( H – J ). P values shown on graphs unless P > 0.05.
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( A ) ELISA for secreted TNFSF13 in colonoid conditioned media ( n = 3 patient lines for control and VEO-IBD; n = 3 passages for variant). ( B ) Representative TNFSF13 RNAscope in colonoids from control and variant participants (scale bar: 50 μm; n = 3 patient lines for control; n = 3 passages for variant). ( C ) Costaining of TNFSF13 and FAS RNAscope probes with Ki67 antibody in colon biopsies. Arrows indicate cells accumulated outside epithelial crypts ( n = 3 patients for control; n = 3 tissue blocks for variant). ( D ) Representative images of colonoid formation assays at day 6 after seeding (scale bar: 300 μm). ( E ) Quantification of newly formed colonoids by size at day 6. Each passage included 2 technical replicates ( n = 4 patient lines for control and VEO-IBD; n = 4 passages for variant). ( F ) TNFSF13 and FAS RNAscope with E-cadherin immunostaining in WT and variant iPSC-derived colon organoids at day 7 (scale bar: 50 μm; n = 3 passages). ( G ) Representative colonoid formation in WT and variant iPSC-derived organoids at day 9 (scale bar: 400 μm; n = 3 passages, each with ≥ 2 technical replicates). ( H ) Quantification of colonoid formation rate and area at day 9. Colonoid size calculated by maximum vertical projection area. ( I and J ) Percentage of EdU + cells following IgG or TNFSF13 neutralizing antibody <t>(nTNFSF13)</t> treatment in ( I ) control tissue–derived colonoids ( n = 3 patient lines) or ( J ) WT iPSC-organoids at day 7 ( n = 3 passages). 2-way ANOVA with multiple comparisons was used for ( A and E ); 2-tailed Student’s t test for ( H – J ). P values shown on graphs unless P > 0.05.
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( A ) ELISA for secreted TNFSF13 in colonoid conditioned media ( n = 3 patient lines for control and VEO-IBD; n = 3 passages for variant). ( B ) Representative TNFSF13 RNAscope in colonoids from control and variant participants (scale bar: 50 μm; n = 3 patient lines for control; n = 3 passages for variant). ( C ) Costaining of TNFSF13 and FAS RNAscope probes with Ki67 antibody in colon biopsies. Arrows indicate cells accumulated outside epithelial crypts ( n = 3 patients for control; n = 3 tissue blocks for variant). ( D ) Representative images of colonoid formation assays at day 6 after seeding (scale bar: 300 μm). ( E ) Quantification of newly formed colonoids by size at day 6. Each passage included 2 technical replicates ( n = 4 patient lines for control and VEO-IBD; n = 4 passages for variant). ( F ) TNFSF13 and FAS RNAscope with E-cadherin immunostaining in WT and variant iPSC-derived colon organoids at day 7 (scale bar: 50 μm; n = 3 passages). ( G ) Representative colonoid formation in WT and variant iPSC-derived organoids at day 9 (scale bar: 400 μm; n = 3 passages, each with ≥ 2 technical replicates). ( H ) Quantification of colonoid formation rate and area at day 9. Colonoid size calculated by maximum vertical projection area. ( I and J ) Percentage of EdU + cells following IgG or TNFSF13 neutralizing antibody <t>(nTNFSF13)</t> treatment in ( I ) control tissue–derived colonoids ( n = 3 patient lines) or ( J ) WT iPSC-organoids at day 7 ( n = 3 passages). 2-way ANOVA with multiple comparisons was used for ( A and E ); 2-tailed Student’s t test for ( H – J ). P values shown on graphs unless P > 0.05.
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Image Search Results


( A ) ELISA for secreted TNFSF13 in colonoid conditioned media ( n = 3 patient lines for control and VEO-IBD; n = 3 passages for variant). ( B ) Representative TNFSF13 RNAscope in colonoids from control and variant participants (scale bar: 50 μm; n = 3 patient lines for control; n = 3 passages for variant). ( C ) Costaining of TNFSF13 and FAS RNAscope probes with Ki67 antibody in colon biopsies. Arrows indicate cells accumulated outside epithelial crypts ( n = 3 patients for control; n = 3 tissue blocks for variant). ( D ) Representative images of colonoid formation assays at day 6 after seeding (scale bar: 300 μm). ( E ) Quantification of newly formed colonoids by size at day 6. Each passage included 2 technical replicates ( n = 4 patient lines for control and VEO-IBD; n = 4 passages for variant). ( F ) TNFSF13 and FAS RNAscope with E-cadherin immunostaining in WT and variant iPSC-derived colon organoids at day 7 (scale bar: 50 μm; n = 3 passages). ( G ) Representative colonoid formation in WT and variant iPSC-derived organoids at day 9 (scale bar: 400 μm; n = 3 passages, each with ≥ 2 technical replicates). ( H ) Quantification of colonoid formation rate and area at day 9. Colonoid size calculated by maximum vertical projection area. ( I and J ) Percentage of EdU + cells following IgG or TNFSF13 neutralizing antibody (nTNFSF13) treatment in ( I ) control tissue–derived colonoids ( n = 3 patient lines) or ( J ) WT iPSC-organoids at day 7 ( n = 3 passages). 2-way ANOVA with multiple comparisons was used for ( A and E ); 2-tailed Student’s t test for ( H – J ). P values shown on graphs unless P > 0.05.

Journal: The Journal of Clinical Investigation

Article Title: TNFSF13 insufficiency disrupts human colonic epithelial cell growth and associated B cell dynamics

doi: 10.1172/JCI186032

Figure Lengend Snippet: ( A ) ELISA for secreted TNFSF13 in colonoid conditioned media ( n = 3 patient lines for control and VEO-IBD; n = 3 passages for variant). ( B ) Representative TNFSF13 RNAscope in colonoids from control and variant participants (scale bar: 50 μm; n = 3 patient lines for control; n = 3 passages for variant). ( C ) Costaining of TNFSF13 and FAS RNAscope probes with Ki67 antibody in colon biopsies. Arrows indicate cells accumulated outside epithelial crypts ( n = 3 patients for control; n = 3 tissue blocks for variant). ( D ) Representative images of colonoid formation assays at day 6 after seeding (scale bar: 300 μm). ( E ) Quantification of newly formed colonoids by size at day 6. Each passage included 2 technical replicates ( n = 4 patient lines for control and VEO-IBD; n = 4 passages for variant). ( F ) TNFSF13 and FAS RNAscope with E-cadherin immunostaining in WT and variant iPSC-derived colon organoids at day 7 (scale bar: 50 μm; n = 3 passages). ( G ) Representative colonoid formation in WT and variant iPSC-derived organoids at day 9 (scale bar: 400 μm; n = 3 passages, each with ≥ 2 technical replicates). ( H ) Quantification of colonoid formation rate and area at day 9. Colonoid size calculated by maximum vertical projection area. ( I and J ) Percentage of EdU + cells following IgG or TNFSF13 neutralizing antibody (nTNFSF13) treatment in ( I ) control tissue–derived colonoids ( n = 3 patient lines) or ( J ) WT iPSC-organoids at day 7 ( n = 3 passages). 2-way ANOVA with multiple comparisons was used for ( A and E ); 2-tailed Student’s t test for ( H – J ). P values shown on graphs unless P > 0.05.

Article Snippet: Meanwhile, to assess the efficiency of neutralizing antibody, nTNFSF13 (Cat #MAB5860, R&D Systems) with a concentration gradient (0, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 20, 30, 50 μg/mL) was added along with 500 ng/mL recombinant Human TNFSF13 protein for each well at 4h post plating.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Variant Assay, RNAscope, Immunostaining, Derivative Assay