GSM9158473: Placebo rep 2; Mus musculus; RNA-Seq
Two mice at 13-weeks-old in each tamoxifen (5-weeks of 5 mg, 60-day release pellet) (Innovative Research of America, SE-361) or placebo pellet (SE-111) inoculated PyMT group were used. Lung tissues were perfused and subsequently inflated via a catheter in the trachea by an enzyme mix containing dispase (2.2 U/mL), collagenase (2 mg/mL), elastase (1 mg/mL), and DNase (5 U/mL). After tying off the trachea, the lung was removed and immediately minced to small pieces. The tissue was transferred into enzyme mix for enzymatic digestion for 30 min at 37°C. After enzyme activity was stopped by adding DMEM/F12 supplemented with 10% FBS, dissociated cells in suspension were passed through a 70 μm strainer and centrifuged at 500 × g for 5 min at 4 °C. The cells were counted using a hematocytometer and assessed for single-cell separation and viability. Single-cell suspensions were processed using the 10x Genomics Chromium Single Cell 5' Gene Expression assay (R2-only chemistry), following the manufacturer's protocol. Barcoded cDNA libraries were generated and sequenced on an Illumina NovaSeq 6000 platform.