GSM6696569: Lw_c02, control, scRNAseq; Mus musculus; RNA-Seq
Liver tissues of mice were obtained from wild-type mice (N=2) and NASH mice with fibrosis (HFD+F+CCl4, N=3; WD+F+CCl4, N=3) and were transferred into a sterile RNase-free tube containing calcium-free and magnesium-free 1x phosphate buffered solution (PBS) on ice. Tissues were transferred into the culture dish and cut it into 0.5 mm 2 pieces,  and washed with 1 × PBS. Liver tissues were dissociated into single cells in dissociation solution 0.35% collagenase IV5,2 mg/ml papain, 120 Units/ml DNase I ) in 37 water bath. The overall cell viability was higher than 85%. Single cell suspensions were loaded to 10x Chromium to capture 5000 single cell according to the manufacturer s instructions of 10X Genomics Chromium Single Cell 3 kit ( V3 ) .The following cDNA amplification and library construction steps were p erformed according to the standard protocol. Libraries were sequenced on an Illumina NovaSeq 6000 sequencing system (paired end multiplexing run,150bp) by LC Bio Technology co.ltd., (HangZhou,China) at a minimum depth of 20,000 reads per cell.