Liver mRNA Sequencing in Mice

After the 2-week treatment, animals were sacrificed for mRNA sequencing from terminal liver samples (Fig 1). RNA was quantified using Qubit (Thermo Fisher Scientific). The RNA quality was determined using a bioanalyzer with RNA 6000 Nano kit (Agilent). RNA sequence libraries were prepared with NeoPrep (Illumina) using Illumina TruSeq stranded mRNA Library kit for NeoPrep and sequenced on the NextSeq 500 (Illumina) with NSQ 500 hi-Output KT v2 (75 CYS, Illumina). Reads were aligned to the Genome Reference Consortium Mouse Build 38 (GRCm38) version 89 Ensembl Mus musculus genome using Spliced Transcripts Alignment to a Reference (STAR) version 2.5.2a with default parameters [27 (link)]. Differential gene expression analysis was carried out using DESeq2 in R programming [28 (link)].