GSM6735360: CD45+ cells, RNA, CTLA4_IDO1 Rep_1; Mus musculus; RNA-Seq
Livers (> 0.5g) were chopped in cold DMEM (Gibco) and digested with the liver dissociation kit, mouse (Miltenyi Biotec) in DMEM according to the manufacturer's instructions. Samples were dissociated with the gentleMACS Octo Dissociator with Heaters (Miltenyi Biotec) as per the default manufacturers protocol for murine liver digestion. The single cell suspension was filtered through a 70 µM mesh strainer and washed with cold DMEM, resuspended in RBC lysis buffer (eBioscience) then washed twice in autoMACs buffer with BSA (Miltenyi Biotec). CD45+ cells were isolated with CD45 microbeads (Miltenyi Biotec) according to the manufacturer's protocol with the autoMACs separator.  The single cell suspension was prepared according to the 10X Genomics single cell sample prep protocol. 6000-8000 cells per sample were loaded onto the 10X Chromium Controller RNA-seq libraries were prepared using Chromium Single Cell 5' v2 Reagent Kits (10X Genomics). libraries were sequenced by UCSD IGM Center on an Illumina NGS HiSeq 4000, with 2x 75 paired-end kits.