GSM5209426: Genome-wide mapping of CTNNB1 binding sites in HepG2; Homo sapiens; ChIP-Seq
TF-DNA complexes were isolated by anti-ASCL2 or anti-CTNNB1. Briefly, the cells were cross-linked with 1% formaldehyde for 10 min at room temperature, and cross-linked cell lysates underwent ultrasonic fragmentation and incubated with antibodies bound to Dynabeads (Thermo Fisher, #10001D and #10003D) at 4°C overnight. The beads were washed and eluted with elution buffer (0.5% SDS, 25 mM Tris-HCl, 5 mM EDTA). The eluates were treated with 1.5 μg of pronase at 42°C for 2 h, then incubated at 65°C overnight to reverse the cross-links. The immunoprecipitated DNA was purified by QIAquick PCR Purification Kit (QIAGEN, #28106). Sequencing libraries were generated using the TruSeq ChIP Sample Prep Kit (Illumina), according to the manufacturer's instructions.