GSM5608855: DKO_LUNG; Mus musculus; RNA-Seq
Eight to 10-week-old female naïve mice of WT, TKO, and DKO genotypes (three mice per group) were perfused with PBS, and lungs were collected. After digestion of lung tissue, percoll was used to enrich immune cells. Briefly, lung samples were suspended in 44% percoll (GE17-0891-01, Sigma Millipore), and then 67% percoll was added slowly and carefully to the bottom of 50 ml tubes. Cells were then spun at 1000 g, 10 mins at 4 °C, and cells at the interface of the percoll gradient were collected and stained with anti-CD45-FITC. Live CD45+ cells were sorted and subjected to single-cell RNAseq. The cell viability was maintained at more than 90% and no less than 10000 cells per group were loaded to generate libraries. RNA libraries were prepared for sequencing using standard Illumina protocols