GSM7956418: Female Knockout non-breeders; Mus musculus; RNA-Seq
No treatments to WT versus KO mice; KO were generated by crossing Tsc2loxP/loxP mice with Tbx4LME_Cre mice. Murine lungs were inflated under constant pressure (25 cm H2O) and the pulmonary circulation was flushed with 2cc of PBS. Lungs were collected and mechanically minced prior to enzymatic digestion with collagenase, dispase and DNase. Samples were then filtered, RBC lysed with ACK lysis buffer and processed into single cell suspension.  CD45+ immune cells were depleted using MACS LS columns with CD45-microbeads with 2 × 10^6 cells per column to enhance purity and viability. Libraries were prepared for sequencing using standard Illumina protocols