title: GSM9251550: shCTR HT1/2/3; Mus musculus; RNA-Seq
library_construction_protocol: The lung lobes were minced on a glass slide and then digested (collagenase IV (Sigma Aldrich C5138), DNAse I (Sigma Aldrich, DN25) in RPMI with 10% FBS) for 35 minutes at 37 degrees celsius. Digestion was stopped with EDTA (1 mM). Digested tissue were filtered into a single cell suspension through a 100 micron filter followed by red blood cell lysis (BD Pharm Lyse 555899). Cells were then washed and suspended in a staining buffer. Cells were then stained with live dead staining (Zombie UV fixabable viability dye, Biolegend #423107) and PeCy7 anti-CD45 (see flow cytometry methods section for staining protocol). Approximately 500,000 lung immune cells from each condition (2 mice per condition) were sorted as live+ IV-CD45 - CD45+ and 50,000 tumor cells were sorted as live+ CD45- GFP+. Followed 10x ECCITE-seq standard library construction protocol.