GSM4727211: MC68NP2; Mus musculus; RNA-Seq
Single cell suspensions were obtained from lung followed by digestion with collagenase IV 0.25mg/ml (Sigma) at 37°C for 30min followed by passing through a 70mm cell strainer and RBC lysis (RBC lysis buffer, BioLegend) for 2min at room temperature. For each mouse scRNAseq sample, 8,500 monocytes and macrophages were sorted as CD45+LINNEG (CD3/CD19/NKP46) CD11B+LY6GNEG and encapsulated using the 10X Chromium 3' v2 chemistry kit according to manufacturer instructions. Sequencing libraries were prepared according to manufacturer instructions. QC of cDNA and final libraries was performed by CyberGreen qPCR library quantification assay (KAPA). Samples were sequenced on an Illumina Nextseq 500 using the 75-cycle kit to a depth of 100 million reads per library.