Fig. 3

Effect of HSF1 depletion on chromatin SUMO2/3 pattern in K562 cells. a Anti-HSF1 antibody immunoblotting of total cellular lysates from shSCR (scrambled control)- or shHSF1-treated cells in C or HS conditions. Anti-tubulin was used as a loading control. b HS-triggered induction of HSP gene expression is blunted in HSF1-silenced (shHSF1) cells. RT-qPCR with primers specific for HSPA1A and HSPB1 was used to measure gene expression. Error bars represent standard deviation of three biological replicates. Stars depict statistical significances between indicated samples (Student’s t-test, **P <0.01, *P <0.05). c HS-induced changes in chromatin SUMO2/3 are generally independent on HSF1 silencing. ChIP-seq tracks of HSF1 in HS, and SUMO2/3 in C and HS conditions in shSCR- and shHSF1-treated cells at indicated chromatin loci. Numbers (on the left) depict maximal signal intensity of each track. d HSF1/SUMO2/3 co-bound loci lose SUMO2/3 signal in HSF1 depleted cells. Two intergenic HS-inducible SUMO2/3 loci with and without HSF1-binding site. Comparison of average SUMO2/3 HS ChIP-seq signals in ±1 kb window at the centers of SUMO2/3 and HSF1 co-occupied peaks and SUMO2/3 peaks without HSF1 in shSCR- or shHSF1-treated cells as indicated