ChIPseq of lung cancer cell IP FOXA3-R1
ChIP was performed using a SimpleChIP Plus Enzymatic Chromatin IP Kit (9003, Cell signaling) according to manufacturers instructions. Briefly, Flag-tagged FOXA3 overexpressed H1299 lung cancer cells were cross-linked by 1% formaldehyde for 10 min and stopped by 0.125 M glycine solution. Cells were then washed twice with ice-cold PBS and scraped into PBS, the extracted chromatin was digested and fragmented into 200 to 1000 bp by the Bioruptor (ScientzIID; Scientz) in SDS lysis buffer, and then immunoprecipitated by ChIP grade antibodies against Flag (F1804, Sigma) or a normal IgG (12-371, Sigma) using Magnetic Beads. After that, the DNA were uncross-linked with proteins.