GSM6652502: WT lung Cd45+ scRNA-seq; Mus musculus; RNA-Seq
Mutant or WT mice were sacrificed 7 weeks after first urethane treatment. Lungs were digested and single cell suspensions were washed with 1XPBS-4%FBS before incubation for 20 min on ice at 5•107 cells/ml with 500 ng/ml Fc block (2.4G2, BD Pharmingen). Cells were then incubated for 1 hour on ice with AF700 conjugated CD45.2 monoclonal antibody (104, Biolegend). For scRNA-seq libraries, DAPI negative (live) CD45+ cells were sorted using a flow cytometer, and sorted cells were resuspended in RPMI for counting. Libraries were prepared using Single Cell 3' Reagent Kits v3.1: ChromiumTM Single Cell 3' Library & Gel Bead Kit v3.1, PN-1000196 and PN-1000129 and following the Single Cell 3' Reagent Kits v3 (PN-1000269) User Guide (Manual Part # CG000315 Rev C). Libraries were sequenced on NovaSeq S4, using half full lanes for 2 samples, with mean read depth of 87-128 thousand reads per cell.