GSM8324092: IP74_3_8_DMSO_H3K27ac_2; Homo sapiens; ChIP-Seq
ChIP-seq experiments were conducted using feeder-free iPSCs, CD45+CD34+ HPCs, and K562 cells. A Dead Cell Removal Kit (Miltenyi Biotec, 130-090-101) was used to eliminate dead cells from feeder-free iPSCs and HPCs. Cells were fixed in PBS with 1% formaldehyde (Thermo Fisher Scientific) with gentle mixing for 10 minutes at room temperature. The reaction was stopped by adding a glycine solution (10x; Cell Signaling Technology) and incubating for 5 minutes at room temperature, followed by two rounds of cold PBS washing. The cells were processed using the SimpleChIP Plus Sonication Chromatin IP Kit (Cell Signaling Technology) according to the manufacturer's protocol. The chromatin was sonicated with Covaris E220 (Covaris) using a microTUBE AFA Fiber Pre-Slit Crimp-Cap (130 µL). Immunoprecipitation of the cross-linked chromatin was performed using antibodies directed against histone H3 lysine 27 acetylation (H3K27ac) (Cell Signaling Technology). ChIP-seq libraries were constructed using a ThruPLEX DNA-Seq Kit (Takara) according to the manufacturer's protocol and sequenced using NextSeq 500 or NovaSeq 6000 (Illumina)