GSM9336626: VSMC, Female, HGPS, P7; Homo sapiens; Hi-C
Briefly, two cell lines of control and HGPS VSMCs (8 × 10^6 cells) were crosslinked in 1% formaldehyde for 10 min at room temperature and quenched with 0.2 M glycine. Crosslinked cells were pelleted by centrifugation and stored at -80 °C. Nuclei were isolated and chromatin was digested overnight with 100 U of DpnII (NEB). Following digestion, 5′ overhangs were filled in with biotin-14-dATP (Life Technologies) and blunt-end ligation was performed in situ for 4h. Crosslinks were reversed by proteinase K treatment, and DNA was purified by phenol:chloroform extraction, ethanol precipitation, and resuspension in low EDTA, tris-buffered water (TLE). For library preparation, 40 µg of DNA from each sample was fragmented to 200–300 bp using a sonicator (Covaris). Fragmented DNA was size-selected with AMPure XP beads (Beckman Coulter) according to the manufacturer's instructions. The purified DNA fragments underwent end repair, after which biotinylated DNA was isolated using Streptavidin beads. Subsequently, polyA tailing and adapter ligation (Illumina TruSeq) were performed to generate the final libraries, and libraries were PCR-amplified and purified.