2.12.1 RNA-seq analysis

For RNA-seq analysis, RNA was extracted using the RNeasy Mini Kit (Qiagen). The quality of isolated RNA was analyzed using the Agilent RNA 6000 Nano Kit (Agilent Technologies, United States), and only RNA samples having RIN > 7.5 and 28S/18S ratio ≥ 0.8 were selected for RNA-seq analysis. Whole transcriptome expression analysis was done using the BGI DNBseq PE100 RNA-seq platform. For data analysis workflow, STAR (Dobin et al., 2013), HTSeq (Anders et al., 2015), and DESeq2 (Love et al., 2014) algorithms were applied. STAR aligner was used to map the reads to the Hg38 reference genome, while HTSeq-count was used for gene quantification and generation of gene expression counts. Differential gene expression analysis was implemented using DESeq2.