A total of 20 million MyPL1 and MyPL2 cells were cross‐linked with 1% formaldehyde (Sigma‐Aldrich, F1635) for 10 min at room temperature, followed by a 5‐min treatment with 0.125 M glycine (Sigma‐Aldrich, G‐8790). Nuclei were isolated and chromatin was purified by chemical lysis. The purified chromatin was fragmented into 200–300 bp fragments by sonication (Covaris, S220) using the truChIP chromatin shearing kit (Covaris). Chromatin immunoprecipitation was performed by incubating the chromatin fraction overnight with 100 μL of protein‐A coated beads (Thermo‐Scientific, 53,139) and 8 μg of the H3K27Ac antibody (Abcam, ab4729). The next day, beads were washed to remove nonspecific binding events and enriched chromatin fragments were eluted from the beads, followed by reverse cross‐linking by incubation at 65°C overnight. DNA was subsequently purified by phenol/chloroform extraction, assisted by phase lock gel tubes (5Prime). DNA obtained from the ChIP assays was adaptor‐ligated, amplified, and analyzed by Illumina sequencing. Raw sequencing data were mapped to the mouse reference genome (GRCm38) using Bowtie. Peak calling was performed using MACS2.
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