HeLa cells were exposed to 44 °C for 1 h and immediately fixed for 10 min with 1% formaldehyde (Carl Roth Inc) at RT followed by 5 min blocking with 125 mM Glycine at RT. Chromatin was extracted using the truChlP Chromatin Shearing Kit (Covaris, #520154) according to the manufacturer´s instructions. The chromatin was sheared by sonication to a DNA fragment size of 200–600 bp and precipitated using an antibody against human BRD4 (Bethyl Laboratories, A301-985A100, RRID:AB_2620184). ChIPs were run on the IP-Star compact system using the Auto iDeal ChIP-seq kit for histones (Diagenode, #C01010051) according to the manufacturer’s recommendations for ChIP preparation. ChIP-DNA was sequenced on a HiSeq4000, 50-bp single-end. Reads were mapped to the hg19 genome using bwa-0.7.12 with default parameters28 (link). Peaks were called with MACS229 (link) with the parameters “bw 500 -mfold 2100 -broad broad-cutoff 0.1 –bdg”. For visualization the bigwig files were converted to bedgraph files that were used for visualization with R/Bioconductor GViz package 30 (link).
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