GSM8604369: cnr.wt.treatment_brm014.ctrl.fra1.rep3; Homo sapiens; OTHER
CUT&RUN was performed following standard CUT&RUN conditions outlined in protocol provided by Henikoff lab on protocols.io (Skene and Henikoff 2017, Janssens 2019, Meers et al. 2019) with minor modification of increasing wash steps to 4 times. Purified pAG-MNase was provided by the NIEHS Protein Expression Core and purified using Meers et al. 2019 protocol. For each reaction, 5e5 cells were harvested. Antibody dilutions (overnight incubation) were as follows: IgG Rabbit (Antibodies online ABIN101961; 1:100), Mouse IgG (Abcam ab46540; 1:100), FLAG M2 (Sigma F1804; 1:50), FRA1 (Abcam ab252421; 1:50), p300 (CST 86377S; 1:50), H3K4me1 (Epicypher 13-0057; 1:50), H3K27ac (Millipore MABE647; 1:50), BRG1 (Abcam ab110641; 1:50), SNF5/SMARCB1 (CST 91735S; 1:50). Library preparations were performed with NEBNext Ultra II (NEB #E7645) described for transcription factors(Liu et al. 2018, Liu 2021) at half volume reactions. Additional modifications were made using NEBNext unique Dual Index Primers (NEB #E6440, #E6442), 15 ng of input DNA, and 0.5 µM diluted adapter.