ChIP-seq Analysis of H3K27ac in Prostate Cancer

ChIP on prostate cancer biopsy and prostatectomy tissues were performed as previously described [61 (link)]. Nuclear lysates of each tissue specimen were incubated with 5 μg of H3K27ac antibody (Active Motif, 39133) pre-bound to 50 μL magnetic protein A Dynabeads (Thermo Fisher Scientific, 10008D). Immunoprecipitated DNA was processed for library preparation (Part# 0801-0303, KAPA biosystems kit), and samples were sequenced using an Illumina HiSeq 2500 system (65 bp, single-end). Sequences were aligned to the human reference genome hg19, duplicate reads were removed, and reads were filtered based on MAPQ quality (≥ 20).