title: GSM8143628: β3-AR-stimulated H2A.Z KD BAs, rep1; Mus musculus; ATAC-seq
library_name: GSM8143628
library_construction_protocol: ATAC-seq was performed using the ATAC-Seq Kit (Active Motif) according to the manufacturer's protocol. Murine brown preadipocytes were seeded in a 6-well plate and differentiated into mature adipocytes. Half an hour prior to performing ATAC-seq in adipocytes treated with vehicle or 1 μM CL316,243 for 4 hours, 200 U/ml DNase I was added to culture medium to eliminate free DNA. Adipocytes on the plate were washed three times with ice-cold PBS, scraped down in 1ml PBS, and counted. A total of 50,000 cells were transferred into a new tube and centrifuged at 500 g for 5 minutes at 4°C. Cell pellets were resuspended with 100 μl ice-cold ATAC lysis buffer and centrifuged at 500 g for 10 minutes at 4°C. Cell pellets were resuspended with 50 μl Tagmentation Master Mix and incubated at 37°C for 30 minutes in a thermomixer set at 800 rpm. The reaction was quenched by adding 250 μl DNA Purification Binding Buffer and 5 μl 3 M sodium acetate. The labeled DNA was purified using DNA purification columns and eluted with 35 μl DNA Purification Elution Buffer. The purified DNA was subjected to library preparation using ATAC-Seq Kit (Active Motif) according to the manufacturer's protocol.