GSM6042897: CAF culture from mouse # 66391 macroH2A1 CUT&RUN prior to serum stimulation; Mus musculus; OTHER
2 × 10^5 short-term cultured CAFs prior to or after 30min serum stimulation were processed for CUT&RUN as described in Skene PJ, Henikoff JG, Henikoff S. Targeted in situ genome-wide profiling with high efficiency for low cell numbers. Nat Protoc. 2018 May;13(5):1006-1019. doi: 10.1038/nprot.2018.015. Epub 2018 Apr 12. PMID: 29651053. Cells were permeabilized with 0.0085% digitonin, incubated with antibody, and DNA cleaved with the CUTANA pAG-MNase (Epicypher). Released DNA was processed using the NEBNext Ultra II Library Preparation Kit for Illumina (NEB), including a 14-cycle amplification step.