title: GSM7903809: S16 TEAD1 CR; Rattus norvegicus; OTHER
library_name: GSM7903809
library_construction_protocol: Plates were washed with PBS containing protease inhibitor cocktail, then washed with nuclear extraction buffer (20mM HEPES pH 7.9, 10mM KCl, 1.5mM MgCl2, 0.5mM Spermidine, 10% Glycerol, 0.3% Triton X-100) to isolate nuclei. Cell nuclei were bound to Concanavalin A beads using the Cut&Run assay kit (Cell Signaling Technology #86652). Nuclei were permeabilized with digitonin, then incubated with protein AG-micrococcal nuclease (PAG MNase). The following antibodies were used for the IP: Nr2f2 (Perseus Proteomics PP-H7147-00), RxRG (Developmental Studies Hybridoma Bank PRCP-RXRG-5H4), and TEAD1 (BD Biosciences 610923). Other antibodies used were: YY1 Active Motif (AM-61980), and H3K27ac  Active Motif (AM-39133), and a SREBP1 antibody provided Tim Osborne PMID: 19666523.  After removal of Concanavalin-bound nuclei, DNA was purified from the remaining solution using the Cell Signaling DNA purification kit (Cat# 14209S). Cut&Run samples were processed using the Illumina NovaSeq6000 system and NEBNext Cut&Run library prep. ChIP-seq was performed as described in Srinivasan et al. PMID: 22492709. NEBNext Cut&Run library prep.