Single-cell RNA sequencing of liver tissues from NASH and control mice

The role of single-cell subsets in NASH is unclear. We used single cell RNA sequencing to analyze the diversity of different single-cell subpopulations in NASH. Male C57BL/6 mice (8 weeks old) were randomly fed either a standard diet (low fat) as a control or a methionine-choline deficient diet (MCD; 40% carbohydrate, 10% fat, deficient in methionine and choline) as NASH mice for 6 weeks (each group, n=3). Liver tissues were cut into about 1-2mm3 pieces and digested in SoloTM Tumor Dissociation Kit (Sinotech Genomics Co. Ltd., Shanghai, China, JZ-SC-58201) at 37°C for 30-60min. Then we stopped enzymatic digestion with excess RPMI-1640 medium, and filtered cell with 40 μm Cell strainer. Single cell solution were kept on ice before loading to BD Rhapsody cartridge for single cell transcriptome capturation. Samples from NASH and control groups were pooled separately for scRNAseq analysis.