title: GSM7830952: STING-/- hypoxia; Mus musculus; RNA-Seq
library_construction_protocol: Mouse whole lungs were perfused with 10mL PBS to remove blood cells. Lung tissues were then cut into small pieces and incubated in 3mL RPMI 1640 with 10% FBS (Gibco #A4766801), 5% Liberase (Sigma Aldrich #05401127001), and 1000U/mL DNAse I (Sigma Aldrich #9003989) for 1h at 100rpm and 37oC. Tissues were triturated with 3mL syringes and 18G needles until complete dissociation. Cells are filtered through 70mm cell strainer, washed 3 times with DPBS 2% FBS and 1M EDTA (KD Medical #RGF-3130) to remove debris. The remaining red blood cells were lysed with ammonium chloride lysis buffer (KD Medical #50-1019080). Single cells were captured in the 10X Genomic Chromium Single Cell 3' Solution, and RNAseq libraries were prepared following manufacturer's protocol. The libraries were subjected to high-throughput sequencing on an Illumina NovaSeq 6000 platform, targeting 6000-8000 cells per sample with a sequencing depth of 20 million reads of 150-bp paired-end reads.