GSM9465548: DEC1 (BHLHE40) ChIP-seq in wild-type bone marrow-derived macrophages (IL-4-treated); Mus musculus; ChIP-Seq
ChIP was performed with minor modifications of the previously described protocol (Daniel et al. Methods Mol Biol. (2014). Briefly, 1x107 BMDMs were double cross-linked with DSG (45 min) and formaldehyde (10 min). Nuclei were isolated using a lysis buffer containing 0.15M NaCl, 0.005M EDTA pH 7.5, 0.05M Tris-HCl pH 7.5, 0.5% NP40, supplemented with protease inhibitor (Roche) prior to use. chromatin was sheared with sonication and immunoprecipitated overnight with the DEC1 antibody (NB100-1800, Novus Biologicals). Chromatin-antibody complexes were extensively washed and eluted after pull-down by paramagnetic beads. Eluted complexes were decrosslinked overnight and purified by the MinElute PCR Purification Kit. Immunoprecipitated DNA was quantified by a Qubit fluorometer. Libraries were prepared from 10ng DNA withTruSeq ChIP library systems (Illumina) according to the manufacturer's instructions. Libraries were sequenced on NextSeq 2000.