Whole Transcriptome Sequencing and Differential Gene Expression Analysis

Whole transcriptome sequencing libraries were constructed using a TruSeq RNA sample preparation v2 kit (Illumina). Sequencing was performed using the 100-bp paired-end mode of the TruSeq Rapid PE Cluster kit and TruSeq Rapid SBS kit (Illumina). The sequencing reads were mapped to the GRCh37.75 human reference genome by using STAR version 2.4.0 26 (link). To quantify gene expression, mapped reads were processed by RSEM version 1.2.18 29 (link). The 'edgeR' method was used to identify differentially expressed genes (DEGs) and the results were filtered on the absolute value of the log 2 (fold change) to more than two 30 (link). We performed gene ontology (GO) analysis using DAVID 31 (link).