Hepatic microRNA signatures in experimental nonalcoholic steatohepatitis

Two-month-old C57BL/6J male mice were placed on chow diet or a diet enriched in high fat, cholesterol, and fructose (Research diet D09100301: 40 kcal% fat, 2% cholesterol, 20 kcal% fructose, HFCF diet) for 1 or 3 months. Liver RNA was isolated and submitted for small RNA sequencing. Total RNA for Illumina sequencing was extracted from mouse liver tissues using a Direct-zol RNA kit. During RNA isolation, DNA was removed by treating the samples with RNase-free DNase to avoid DNA contamination. Three biological replicates in each group for a total of 12 samples were submitted for RNA sequencing. cDNA libraries were prepared using the Illumina TruSeq stranded RNA kit with Ribo-Zero Gold. Fifty-cycle single-read sequencing was performed with an Illumina HiSeq 2500.