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Chromatin immunoprecipitation (ChIP) assay

CM Cunying Ma
XW Xiaoying Wang
FY Fenghua Yang
YZ Yichen Zang
JL Jiansong Liu
XW Xinyi Wang
XX Xia Xu
WL Wenjuan Li
JJ Jihui Jia
ZL Zhifang Liu
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Treated GC cells were cross-linked in 1% (vol/vol) formaldehyde-containing medium for 10 min at 37 °C and then lysed and sonicated to disrupt chromatin DNA into fragments between 200 and 1000 bp. ChIP was performed using the ChIP Assay 17–295 kit (Merck Millipore, Germany) according to the manufacturer’s protocol. An antibody against Flag (Sigma-Aldrich, USA) was used to immunoprecipitate the DNA fragments. The protein-DNA complexes were collected with protein A Sepharose beads, eluted and reverse cross-linked. The samples were extracted with Dr.GenTLE™ Precipitation Carrier (TaKaRa Biotechnology Co., Ltd., Dalian, China). The recovered DNA was resuspended in DDW and used as the template to amplify the ADAR1 promoter. The primer sequences are listed in Table S3.

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