GSM8239766: THP-1 cell line, SPI1 knockdown, H3K27ac, ChIP-seq, rep1; Homo sapiens; ChIP-Seq
After the cells were transinfection for 48 hours, 10000000 cells were used per immunoprecipitation. At first, the cells were fixed with 1% formaldehyde (Sigma-Aldrich, 252549) at room temperature for 10 min. A final concentration of 0.2 M glycine was added to quench the fixation. Nuclear lysates were fragmented by Covaris S220 (Adaptive Focused Acoustics®). For each tissue, 3 μg of H3K27ac (Abcam Cat# ab177178) antibodies were mixed with 25 μL Protein A/G Dynabeads (Selleck. cn, B23201), with rotation at 4°C for 4 h. For per immunoprecipitation, about 50 μg of chromatin was incubated with antibody–bead complexes overnight at 4 °C. Chip-DNA was purificated by MinElute PCR Purification kit (QIAGEN, Cat# 28004 ). ChIP-seq libraries were constructed using the MGIEasy DNA library prep kit (1000006985, BGI-Shenzhen, China), and sequenced as 100 bp paired-end reads with the DNBSEQ (MGI-Shenzhen, China) platform.