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yes1 antibody  (Proteintech)


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    Structured Review

    Proteintech yes1 antibody
    ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and <t>YES1.</t> ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.
    Yes1 Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/yes1+antibody/pmc12662217-263-89-91?v=Proteintech
    Average 93 stars, based on 7 article reviews
    yes1 antibody - by Bioz Stars, 2026-07
    93/100 stars

    Images

    1) Product Images from "FeaSion decodes the regulatory landscape and functional diversity of RNA polymerase II CTD phosphorylation"

    Article Title: FeaSion decodes the regulatory landscape and functional diversity of RNA polymerase II CTD phosphorylation

    Journal: Science Advances

    doi: 10.1126/sciadv.adz2345

    ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and YES1. ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.
    Figure Legend Snippet: ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and YES1. ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.

    Techniques Used: CRISPR, Western Blot, Control, ChIP-sequencing, In Vitro, Phospho-proteomics, Activity Assay, FLAG-tag, Binding Assay, Knock-Out



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    ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and <t>YES1.</t> ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.
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    ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and <t>YES1.</t> ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.
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    ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and <t>YES1.</t> ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.
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    ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and <t>YES1.</t> ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.
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    ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and <t>YES1.</t> ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.
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    ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and <t>YES1.</t> ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.
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    Image Search Results


    Up and down-regulation of all significantly changed organ damage-related proteins during HOPE. ( A ) SMAD1; mothers against decapentaplegic homolog 1, ( B ) ALDH3A1; aldehyde dehydrogenase, dimeric NADP-preferring, ( C ) TOP2B; DNA topoisomerase 2-beta, ( D ) PGF; placenta growth factor, ( E ) ENAH; protein enabled homolog, ( F ) FES; tyrosine-protein kinase Fes/Fps, ( G ) LTA4H; leukotriene A-4 hydrolase, ( H ) FOXO1; forkhead box protein O1, ( I ) CA12; carbonic anhydrase 12, ( J ) PLIN1; perilipin-1, ( K ) YES1; tyrosine-protein kinase Yes, ( L ) MAEA; macrophage erythroblast attacher, ( M ) MAGED1; melanoma-associated antigen D1, ( N ) MAX; protein max, ( O ) FOSB; protein fosB. *, non-adjusted p value of less than 0.05; HOPE, hypothermic machine perfusion; NPX, normalized protein expression; T, time in minutes.

    Journal: Asaio Journal

    Article Title: Candidate Biomarkers YES1, Troponin I, Lactate, and Ammonia for Evaluation of Cardiac Function Post Hypothermic Oxygenated Perfusion

    doi: 10.1097/MAT.0000000000002419

    Figure Lengend Snippet: Up and down-regulation of all significantly changed organ damage-related proteins during HOPE. ( A ) SMAD1; mothers against decapentaplegic homolog 1, ( B ) ALDH3A1; aldehyde dehydrogenase, dimeric NADP-preferring, ( C ) TOP2B; DNA topoisomerase 2-beta, ( D ) PGF; placenta growth factor, ( E ) ENAH; protein enabled homolog, ( F ) FES; tyrosine-protein kinase Fes/Fps, ( G ) LTA4H; leukotriene A-4 hydrolase, ( H ) FOXO1; forkhead box protein O1, ( I ) CA12; carbonic anhydrase 12, ( J ) PLIN1; perilipin-1, ( K ) YES1; tyrosine-protein kinase Yes, ( L ) MAEA; macrophage erythroblast attacher, ( M ) MAGED1; melanoma-associated antigen D1, ( N ) MAX; protein max, ( O ) FOSB; protein fosB. *, non-adjusted p value of less than 0.05; HOPE, hypothermic machine perfusion; NPX, normalized protein expression; T, time in minutes.

    Article Snippet: YES1 (BS-4166R, Bioss antibodies, Woburn, MA) immunohistochemistry (IHC) was performed on 4 μm tissue slides.

    Techniques: Expressing

    YES1 antibody-positive stained cardiac tissue during hypothermic and normothermic machine perfusion over time. Representative YES1 immunohistochemistry images of three porcine hearts ( A–J ) depict a heterogeneous staining pattern. At baseline (T0; A,D,G ), after 4 hours of HOPE (T4; B,E,H ) and at the end of 4 hours NMP (T240; C,F,J ), YES1 is localized in the cytoplasm of endothelial cells and cardiomyocytes with varying intensity. The tissue architecture in these slides remains intact throughout HOPE and NMP.

    Journal: Asaio Journal

    Article Title: Candidate Biomarkers YES1, Troponin I, Lactate, and Ammonia for Evaluation of Cardiac Function Post Hypothermic Oxygenated Perfusion

    doi: 10.1097/MAT.0000000000002419

    Figure Lengend Snippet: YES1 antibody-positive stained cardiac tissue during hypothermic and normothermic machine perfusion over time. Representative YES1 immunohistochemistry images of three porcine hearts ( A–J ) depict a heterogeneous staining pattern. At baseline (T0; A,D,G ), after 4 hours of HOPE (T4; B,E,H ) and at the end of 4 hours NMP (T240; C,F,J ), YES1 is localized in the cytoplasm of endothelial cells and cardiomyocytes with varying intensity. The tissue architecture in these slides remains intact throughout HOPE and NMP.

    Article Snippet: YES1 (BS-4166R, Bioss antibodies, Woburn, MA) immunohistochemistry (IHC) was performed on 4 μm tissue slides.

    Techniques: Staining, Immunohistochemistry

    ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and YES1. ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.

    Journal: Science Advances

    Article Title: FeaSion decodes the regulatory landscape and functional diversity of RNA polymerase II CTD phosphorylation

    doi: 10.1126/sciadv.adz2345

    Figure Lengend Snippet: ( A ) Stacked bar plot showing the kinases (left) and phosphatases (right) identified in RNAPII ChIP-MS, CRISPR screening and RNAPII mut ChIP-MS. ( B ) Pie plots showing the family of kinases identified in CRISPR screening. ( C ) Heatmap showing the NormZ of CRISPR screening for CLK1, CLK4, and YES1. ( D ) Western blot analyses of the RNAPII, pY1, pS2, pT4, pS5, and pS7 in CLK1, CLK4, or YES1 KO cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as the loading control. ( E ) Metagene profiles and boxplots (top) and log 2 (fold change) metagene heatmaps (bottom) showing pS7 and RNAPII ChIP-seq density in control, CLK1 KO, and CLK4 KO cells, as well as in untreated and TG003-treated (20 μM, 4 hours) cells (top). Statistical significance was assessed by a two-sided Wilcoxon test. *** P < 0.001. The value on the box plot indicating the fold change. ( F ) Similar to (E), but for pY1 and RNAPII ChIP-seq in control and YES1 KO cells, as well as in untreated and CHR6953755-treated (10 μM, 24 hours) cells. ( G ) Schematic workflow of in vitro phosphorylation assay. ( H ) In vitro phosphorylation activity of CLK1 (left), CLK4 (middle), and YES1 (right) on CTD. The CTD was detected using HA tag, and CLK1, CLK4, and YES1 were detected using Flag tag. ( I ) Pie chart (top) and metagene heatmap (bottom) showing the distribution of the top 500 CLK1 (left) CLK4 (middle) and YES1 (right) ChIP-seq binding sites and their densities. ( J ) Bar plot showing the gene ontology enrichment result of CLK1 (top), CLK4 (middle) and YES1 (bottom) binding genes. ( K ) Tracks of ING5 of CLK1 (left), DLX2 of CLK4 (middle), and IMPACT of YES1 (right) ChIP-seq signals and the changes of pS7 or pY1 and RNAPII after their knockout. miRNA, microRNA; snoRNA, small nucleolar RNA. TES, transcription end site.

    Article Snippet: The membrane was blocked with 5% nonfat dry milk at room temperature for 30 min and then incubated overnight at 4°C with the RPB1 NTD antibody (Cell Signaling Technology, #14958), phospho-RPB1 CTD (Ser 2 ) antibody (Cell Signaling Technology, #13499), phospho-RPB1 CTD (Ser 5 ) antibody (Cell Signaling Technology, #13523), phospho-RPB1 CTD (Tyr 1 ) antibody (Merck Millipore, #MABE350), phospho-RPB1 CTD (Thr 4 ) antibody (Cell Signaling Technology, #26319), phospho-RPB1 CTD (Ser 7 ) antibody (Cell Signaling Technology, #13780), CLK1 antibody (Santa Cruz Biotechnology, #sc-515897), CLK4 antibody (Proteintech, #31205-1-AP), YES1 antibody (Proteintech, #20243-1-AP), α-tubulin antibody (Proteintech, #66031-1-Ig), GFP antibody (Proteintech, #50430-2-AP), HA antibody (Proteintech, #51064-2-AP), or glyceraldehyde-3-phosphate dehydrogenase antibody (Proteintech, #10494-1-AP) to the target protein.

    Techniques: CRISPR, Western Blot, Control, ChIP-sequencing, In Vitro, Phospho-proteomics, Activity Assay, FLAG-tag, Binding Assay, Knock-Out