GSM8972279: WT HeLa, BMN673 treated; Homo sapiens; ChIP-Seq
Treated HeLa cells were cross-linked with 1% formaldehyde for 10 min at room temperature, quenched with 125 mM glycine, and washed with ice-cold PBS. Cells were lysed in SDS lysis buffer (1% SDS, 10 mM EDTA, 50 mM Tris-HCl pH 8.1) supplemented with protease inhibitors, and chromatin was sheared to 200–500 bp fragments using a Covaris S220 sonicator (peak power: 75 W, duty factor: 20%, cycles/burst: 200, duration: 15 min). Sheared chromatin was diluted 10-fold in ChIP dilution buffer (0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-HCl pH 8.1, 167 mM NaCl) and pre-cleared with Protein G magnetic beads (Thermo Fisher, #88847) for 1 h at 4°C. Immunoprecipitation was performed overnight at 4°C using 10 µl of STING antibody (Abcam, #ab239074). Antibody-chromatin complexes were captured with Protein G beads, washed sequentially with low-salt, high-salt, LiCl, and TE buffers, and eluted in elution buffer (1% SDS, 0.1 M NaHCO3). Cross-links were reversed by incubating at 65°C overnight with 200 mM NaCl. DNA was purified using the QIAquick PCR Purification Kit (Qiagen, #28104). Libraries were prepared with the NEBNext Ultra II DNA Library Prep Kit (NEB, #E7645S) following manufacturer instructions.
GEO
GSM8972279_r1