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.
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