RNA Sequencing Data Analysis

For RNA sequencing, total RNA was extracted from liver or cell samples using TRIzol reagent (T9424; Sigma-Aldrich; St. Louis, MO, USA) as indicated above. Then 200 ng of RNA input per sample and a MGIEasy RNA Library Prep Kit (1000006383; MGI Tech; Shenzhen, China) were used to construct cDNA libraries, according to the manufacturer’s instructions. Single-end libraries were sequenced using MGISEQ 2000 (MGI Tech; Shenzhen, China). For data processing, HISAT2 software (version 2.1.0)^{54 (link)} was used to map the sequences from clean reads to Ensembl mouse (mm10/GRCm38) reference genomes. SAMtools software (version 1.4)^{55 (link)} was used to sort and convert the mapped reads to the Binary Alignment Map (BAM) files. StringTie software (version 1.3.3b)^{56 (link)} was used to calculate the fragments per kilobase of exon model per million mapped fragments (FPKM) values of each gene. DESeq2 software (version 1.2.10)^{57 (link)} was used to analyze differential gene expression. Genes with a fold change greater than 1.5 and corresponding adjusted p values less than 0.05 were identified as DEGs.