RNA-Seq of adult rat female liver
RNA reagents were used to extract the total RNA from 20 samples of Sprague-Dawley rats used in this study, according to the manufacturer's protocol (HUAYUEYANG Biotechnology Co. Ltd, Beijing, China). The purity of the RNA samples was assessed at A260/230 nm and A260/280 nm ratios using NanoDrop 2100 (Thermo Fisher Scientific, Waltham, MA, USA). The integrity of the RNA samples was determined using the RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA), and the RNA concentration and genomic DNA contamination were determined using the Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA, USA). The 260/280 ratio of all samples varies from 1.8 to 2.0, and integrity number (RIN) values are greater than 7.0, indicating good RNA quality [38]. A total of 3 g RNA from each sample was used to prepare the cDNA libraries using the NEBNext UltraTM Directional RNA Library Prep Kit from Illumina (NEB, San Diego, CA, USA), following the manufacturer's recommendation. Paired-end (2 x 150 pb) reads of samples in H30 and H60 were sequenced using the HiSeq 2500 sequencer (Illumina, San Diego, CA, USA).