Total RNA was extracted using RNeasy Mini Kit (Qiagen) according to vender's protocol. RNAs were retrotranscribed to cDNA using iScript Supermix™ (Bio-Rad). qPCR was performed using TaqMan. Gene Expression Master Mix (Thermo Scientific) mRNA levels were normalized by comparison to GAPDH mRNA. 100ng total RNA was extracted via polyA-tail specific protocol according to Illumina protocol. Sequencing libraries were generated using the TruSeq stranded RNA sample prep kit (Illumina). The libraries were sequenced as 75 bp single-end reads using an Illumina NovaSeq SP platform with an average of 35M reads per sample. Indexed libraries that met appropriate cut-offs for both quantity and quality were quantified by qRT-PCR using a commercially available kit (KAPA Biosystems) and insert size distribution was determined with the LabChip GX or Agilent Bioanalyzer. Samples with a yield of ≥0.6 ng/ul were used for sequencing.