GSM7153078
Whole right lung superior lobe was isolated from each animal, mouse lung dissociation kit (Miltenyi Biotec) was used with some additional modifications. Following single cell preparation, cell suspensions were applied to a MACS Smart Strainer (70 μm) and washed twice with 10 ml of DMEM. Cells were pelleted by spinning at 300 × g for a total of 7 minutes at 4°C. Cells were resuspended in 1ml DMEM containing 200 μl DNase (1μg/μl) for 5 minutes at room temperature. Cells were washed by adding 10 ml sterile PBS containing 0.5% BSA and pelleted at 4°C. The RBC lysis was carried out using 1X RBC lysis solution (Miltenyi Biotec; 130-094-183) in a total volume of 1ml for 10 minutes at 4°C. Cells were resuspended in 10 ml chilled DPBS containing 0.5% BSA and pelleted subsequently. Cells were filtered using 35 μm Falcon cell strainer and transferred to a 2 ml lo-binding tube in a total volume of 1ml DPBS containing 0.04% BSA. Chromium Next GEM Single Cell 3′ Reagent Kits v3.1 (Dual Index) (CG000315).