Acetyl histone h3 lys27 h3k27ac

ChIP assays were performed using the Magna ChIP A-Chromatin Immunoprecipitation Kit according to the manufacturer's instruction (Cat. 17610, Millipore, USA). The antibodies for Histone H3 (Cat. ab1791) and H3 trimethyl Lys4 (H3K4me3, Cat. ab8580) were from Abcam, and acetyl-histone H3 Lys27 (H3K27Ac, Cat. 4353T) was from Cell Signaling Technology. The ChIP primers were listed in Table. S1. Quantification of immunoprecipitated DNA was performed using qPCR. ChIP data was calculated as a percentage relative to the input DNA from the equation 2[Input Ct − Target Ct] × 100 (%).

ChIP experiments were conducted as previously described [22 (link)]. Briefly, 2 × 10⁶ cells were crosslinked with 1% formaldehyde for 10 min at room temperature. Following cell lysis, the cells were sonicated by a Bioruptor (Diagenode) for 30 min to obtain fragmented DNA. ChIP was incubated with the appropriate antibodies against mono-methyl-histone H3 (Lys4) (H3K4me1, #5326), acetyl-histone H3 (Lys27) (H3K27ac, #8173), and FLAG (#14793) from Cell Signaling Technology (Shanghai, China), followed by a pull-down assay using protein A/G-conjugated beads. The collected DNA was then used for quantitative PCR (qPCR) analysis. The primer sets used are listed in the Supplementary Table.