title: scRNA-seq of Mus musculus: 4 days old liver
design: Single hepatic immune cells were isolated and sorted (n=3) of 4 days old untreated SPF mice. Cells from each mouse were hashtagged with Feature Barcoding technology (TotalSeq C #1-#3, Biolegend). Cell sorting was performed on a FACS ARIA II (BD) equipped with a 100 mm nozzle for live CD45+ cells and using the BD FACS DIVA 8.0.1 software. Liver immune cells were further processed using the Chromium Next GEM Single Cell 3 Reagent Kits v3.1 according to manufacturers instruction (10x Genomics, Inc., United States). 10,000 single immune cells were aimed for sequencing. Single-cell sequencing on High-Output flow cells using 150 cycles was performed on an Illumina NextSeq 500 platform.