ChIP-seq
A 1% formaldehyde solution is introduced into the cytoplasm for cross-linking. Crosslink the solution for 7 minutes. After that, the CHIP proceed with ChIP using the ChIP Assay Kit (E-GENE, China). Add lysis buffer (200 l lysis buffer without additives + 4 l of 50 protease inhibitor cocktail) and incubate on ice for 5 minutes to lyse the cells. Use a Bioruptor sonicator to fragment the DNA into pieces sized between 250 and 750 bp. After centrifugation, collect the supernatant, save a portion as Input, and use the remainder for immunoprecipitation (IP), maintaining a ratio of Input to IP of 10:1. Wash protein A/G magnetic beads (Invitrogen, USA) with PBS supplemented with BSA, then resuspend the beads in the same buffer. Incubate the IP-grade antibody with the beads at 4C for 6 hours before proceeding with washing. Finally, resuspend the beads in dilution buffer and incubate overnight at 4C to capture the DNA-protein complex. Use wash buffer to eliminate any non-specific DNA-protein interactions. To decrosslinking the antibody-enriched DNA-protein complex, incubate with decrosslinking solution at 65C for 3 hours. After this incubation, collect the supernatant and recover the DNA. Measure the DNA concentration using the Qubit dsDNA HS Assay Kit (Invitrogen, USA) in Qubit  2.0 Flurometer. Library construction is performed by E-GENE Biotech Inc. (Shenzhen, China), and it was quantitatively and qualitatively detected using the Qubit dsDNA HS Assay Kit and Agilent 2100 bioanalyzer. Finally, paired-end sequencing is conducted on the Illumina HiSeq PE150 platform (Illumina, San Diego, CA, USA).