title: GSM8433844: TA33L.Combo; Mus musculus; RNA-Seq
library_construction_protocol: Cells were sorted with the Flow cytometry (FC) before sequencing. Organs were smashed into small pieces and then incubated 15min at 37°C in agitation with a tissue digestion solution composed of 1 mL IMDM (Corning) supplemented with 0.35 mg/mL collagenase type IV (Sigma-Aldrich/ SCR103), 1 mg/ml dispase II (Gibco/ 17105041) and 0.2 mg/ml DNAse (Roche/ 11284932001). The tissue was then further dissociated by pipetting and filtered using 40 μm cell strainers (Corning/ 352340). Single cell solution was washed with 30 mL of autoMACS Running Buffer (Miltenyi Biotec/130-940 091-221) and pelleted for 5min 300g 4°C to remove excess enzymes. Samples were transferred to FACS tubes, stained with surface antibody and hashtags for sample identification. Then CD45+ cells were sorted. Library was constructed using a microfluidics-based approach on Chromium Controller (10X Genomics) using the Chromium Next-GEM Single Cell 3' Reagent Kit v2 (10X Genomics) according to the manufacturer's instructions. For each mouse, 30,000 CD45+ cells were sorted.  We then loaded 20,000 cells belonging to 3 mice from the same group onto the Chromium Controller (10X Genomics).