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Fig. 5 | Genome Biology

Fig. 5

From: Global SUMOylation on active chromatin is an acute heat stress response restricting transcription

Fig. 5

Chromatin-bound PIAS1 promotes SUMOylation on VCaP cell chromatin. a Total cellular PIAS1 levels do not change in 30 min HS at 43 °C (HS) or 1 h recovery at 37 °C (Re), as judged by immunoblotting of total cellular lysates with anti-PIAS1 antibody. Anti-tubulin antibody was used as a loading control. b Genomic location of SUMO2/3 peaks in each condition in annotated chromatin loci. c Venn-diagrams showing overlap of PIAS and SUMO2/3 peaks in control (C), HS (HS), and recovery (Re) conditions. d Heat map showing PIAS1 and SUMO2/3 ChIP-seq signal intensities in control (C), HS, and recovery (Re), using false-color scale in ±2 kb windows centered at PIAS1 binding sites. Blue bar depicts PIAS1 peak cluster in control conditions, red bars clusters in HS, and orange bars in recovery. e Depletion of PIAS1 with specific siRNA (siPIAS1) as confirmed by immunoblotting (siNON, non-targeting control siRNA). Anti-histone 3 (H3) was used as a loading control. f SUMO2/3 ChIP-seq signal was reduced at PIAS1 co-occurring chromatin sites, such as the promoter of HS-inducible UBB, in PIAS1-depleted cells. ChIP-seq signals in C and HS conditions are marked with blue and red, respectively. g Heat map showing SUMO2/3 ChIP-seq signal in siNON- and siPIAS1-transfected cells in C and HS conditions at ±2 kb window centered at PIAS1 binding. ChIP-seq signal intensities are shown as false-color. Bars represent PIAS1-binding sites in C (blue) and HS (red) conditions. h Comparison of average HS SUMO2/3 ChIP-seq signals at SUMO2/3 peaks with or without co-occurring PIAS1 in siNON- (red line) and siPIAS1- (dark red dotted line) transfected cells

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