GSM5573045: CK_6_weeks; Mus musculus; RNA-Seq
Lungs were collected from two CK 6w and 6 CC (control) mice injected with 4 doses of tamoxifen. Tissues were dissociated and a single cell suspensions were subjected to immune cell depletion using CD45 microbeads as described previously. fGFP+ cells were enriched by performing FACS in “yield-sort” mode. This mode is suitable to enrich populations found in small numbers, however, also results in contamination from other fGFP- cell types. The number of fGFP+ cells in each lung is small and a minimum of 100,000 cells are necessary for scRNA-seq, hence the 'yield-sort' mode was chosen. fGFP+ cell enriched suspensions from all controls were pooled but the two tumour samples were handled separately. Cells were then resuspended in 0.01% BSA in PBS. scRNA-seq was performed using Drop-seq protocol (Macosko et al., 2015) with modifications described in (Kobayashi et al., 2020). Libraries were sequenced on HiSeq in 150 bp PE configuration.