GSM8436413: PB2_TGFb1_IgG; Homo sapiens; OTHER
Briefly,~500,000 untreated and TGF-b treated NK cells were immobilized to activated Concanavalin A-coated magnetic beads (Bangs laboratories), then permeabilized with wash buffer containing 0.05% digitonin. Samples were incubated with rabbit IgG (Millipore) or BATF (Cell Signaling Technologies) overnight at 4°C. Targeted chromatin digestion was achieved by pAG-MNase (EpiCypher) binding followed by incubation with CaCl2. CUT&RUN DNA fragments were purified using MinElute columns (Qiagen) and libraries were created using NEBNext Ultra II DNA Library prep kit (New England Biolabs) following modified protocol for BATF CUT&RUN DNA as previously described. Libraries were sequenced using Illumina NextSeq 500 system to obtain 75bp paired end reads per sample.