GSM9287926: B6, lung mesenchymal, P8, ATAC-seq, replicate 1; Mus musculus; ATAC-seq
The lungs were perfused with 12 ml of cold DPBS (Life Technology) and then inflated with dissociation buffer (PRMI1640 (Thermo Scientific) with 10% FBS, 1mM HEPES (Life Technology), 1mM MgCl2(Life Technology), 1mM CaCl2 (Sigma-Aldrich), 0.525mg/ml collagenase D (Roche), 5unit/ml Dispase (Stemcell Technologies) and 0.05 mg/ml DNase I (Roche)). Minced lung was placed in dissociation buffer for 30 min at 37°C, and cell solution was filtered through 70μm and 40μm strainers. Ammonium-Chloride-Potassium (ACK) lysis was used to remove blood cells, and cell pellets were resuspended in FACS buffer. Antibodies used for flow cytometry include BV510-CD45, PE-Epcam, PE-CD31, APC-PDGFRa (all from BioLegend), and DAPI to label dead cells. 18,000 cells from each group were subjected to scATAC-seq following standard protocol by Center for Epigenomics at the University of California, San Diego. Sample preparation and library construction were performed according published protocol doi:10.7554/eLife.62522