GSM6625213: Th2_D1_K27Ac_141104_0303_s20; Mus musculus; ChIP-Seq
cell isolation protocol: Cells from spleen were obtained by mechanical disruption. Peritoneal cells were obtained by  washing of the peritoneal cavity with cold PBS. Isolated cells were further sorted as based on their surface markers. in vitro culture protocol: All cells were stimulated in RPMI medium with 10% (vol/vol) FCS (Invitrogen), 2 mM glutamine (Invitrogen), 100 IU/ml penicillin (Invitrogen), 0.1 mg/ml streptomycin (Invitrogen), and 20 mM HEPES buffer, pH 7.2–7.5 (Invitrogen), and 2 mM β-mercaptoethanol (Sigma-Aldrich). NK cells were treated with 1000 U/ml IL-2 and 10  ng/ml IL-12 (R&D) for 6 hours; ILC2 cells were treated with 50 ng/ml IL-25 (BioLegend) and 50 ng/ml IL-33 (Biolegend) for 4 hours; NCR+ILC3 were treated with 50ug/ml of IL-23 (R&D Systems) for 6 hours. ChIP was performed for transcription factors CTCF (Millipore Sigma, #07-729), Rad21 (Abcam, #ab992) and p300 (Abcam, #ab14984) using SimpleChIP Plus Enzymatic Chromatin IP Kit (Cell Signaling Technology, #9005S). ChIP was performed for histone marks H3K27ac (Abcam, #ab4729), H3K27me3 (Thermo Fisher Scientific, #MA511198) and H3K4me1 (Abcam, #ab8895) using iDeal ChIP-seq Kit for Histones (Diagenode, #C01010051). The purified enriched DNA was used to generate libraries using the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, #E7645) and NEBNext Multiplex Oligos for Illumina (NEB, #E6609).