GSM5702452: Mouse Liver; Mus musculus; RNA-Seq
Splenocytes and thymocytes were harvested by mechanical disruption on 70 μm nylon mesh followed by red blood cell (RBC) lysis and washing with Hank's Balanced Salt Solution (HBSS) (Gibco) supplemented with 10% FBS. Lung tissue was digested with Stemcell spleen dissociation medium, and mechanically dissociated using GentleMacs Dissociator (Miltenyi). Cells were strained though a 70 μm filter and washed with HBSS supplemented with 10% FBS followed by RBC lysis. Liver cells were harvested by mechanical disruption on 70 μm nylon mesh followed by 34% Percoll gradient before RBC lysis and washing. For MAIT cell enrichment before sorting, negative selection of cells was carried out using biotinylated antibodies against CD11b (clone M1/70), CD11c (clone M418), F4/80 (clone BM8.1), CD19 (clone 1D3), and TER-119 (clone TER-119).  These were used together with Rapidspheres (Stemcell Technologies) and either the Big Easy (Stemcell Technologies) or Easy eight magnets (Stemcell Technologies) and protocols from Stem Cell Technologies. MAIT cells were sorted using a FACSAria III (BD Biosciences). Single-cell libraries prepared as per the manufacturer's instructions (10x Genomics). Samples were processed using 10X v2 chemistry, as per the manufacturer's recommendations.