RNA-seq Library Preparation and Analysis

Total RNA was extracted from each cell group using Trizol reagent (Thermo Fisher Scientific, Waltham, MA, United States) according to the manufacturer’s instructions. RNA sample quality was assessed using Nanodrop 2000 (ThermoFisher) and BioAnalyzer 2100 (Aglient Technologies). The RNA-seq libraries were sequenced using a TruSeq Stranded mRNA library preparation kit [(Illumina, San Diego, CA, United States)] and constructed as well as sequenced using HiSeq × Ten (Illumina) by BioMarker (Beijing, China). The RNA-seq reads were trimmed, filtered, and quality-controlled using FastQC (Babraham Institute) tools. Reads per kilobase per million mapped reads (RPKM) were then calculated using Hisat2 (Kim et al., 2015 (link)).