GSM8464064: Cal27, BRD4, ChIP; Homo sapiens; ChIP-Seq
Cal27 cells (~1×107) in a logarithmic growth phase with monolayer growth were washed and collected. All samples were fixed with 1% formaldehyde at room temperature for 15 min, followed by 0.125 M glycine treatment for 5 min. The sample was then pelleted, washed and re-suspended in ChIP lysis buffer (25 mM Tris-HCl pH 7.5, 1 mM EDTA, 500 mM NaCl, 1% Triton X-100, 0.1% sodium deoxycholate, 0.05% SDS, 1×Proteinase Inhibitor Cocktail). Then, samples were sonicated by Bioruptor® Plus sonication device (Shearing condition: 20 cycles on cooling condition, 30 seconds ON, 30 seconds OFF), immunoprecipitated with antibody against H3K27ac, BRD4, EP300, SOX2, SMAD3 or Rabbit IgG on ChIP-Grade Protein G Agarose Beads (#9007, Cell Signaling Technology, USA) and purified by Universal DNA Purification Kit (#DP214, TIANGEN Biotech, China). The ChIP-Seq library was prepared using NEBNext Ultra II FS DNA Library Prep Kit for Illumina (#E7645L, New England Biolabs, USA) according to the manufacturer's instructions.