RNA sequencing analyses
Total RNA was extracted using the mirVana miRNA Isolation Kit (Ambion)
following the manufacturer’s protocol. The integrity of RNA was assessed
using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara,
CA, USA). Samples with an RNA integrity index greater than 7 were used
in subsequent analyses. Libraries were constructed using the TruSeq
Stranded mRNA LT Sample Prep Kit (Illumina, San Diego, CA, USA)
following the manufacturer’s protocol. Sequencing of these libraries was
then conducted on an Illumina sequencing platform (HiSeqTM 2500 or
Illumina HiSeq X Ten), and 125 bp/150 bp paired-end reads were
generated.