ChIP-qPCR assay

Indicated ICC cells were treated with 200 nM of F-A Cho-HDO for 72 h before they were subjected to chromatin preparation for the ChIP analysis. ChIP assays were conducted using the Simple ChIP Enzymatic Chromatin IP Kit (Cell Signaling Technology, Danvers, MA, USA) according to the manufacturer’s instructions. Briefly, ICC cells with indicated treatments were crosslinked with 1% formaldehyde for 20 min and terminated by addition of glycine (150 mM). Then cells were lysed, and chromatins were sonicated to an average length of 200–1,000 base pairs. One hundred microliters of chromatins were immunoprecipitated overnight at 4°C using 10 μg of anti-STAT1, non-specific anti-immunoglobulin G was served as the negative control. Then 20 μL of protein A beads was added and incubated for 2 h. After reverse cross-linking and DNA purification, immunoprecipitated DNA was quantified by RT-qPCR. The primers used for RT-qPCR analysis of precipitated DNA are listed in Table S6.