350 ul QIAGEN Buffer RLT plus was added to dissolve the thawed cell pellets. The QIAGEN RNeasy Plus mini (QIAGEN, Cat. #74134) kit was used for RNA purification on QIAcube automation (QIAGEN, Cat.# 9002864). RNA samples were eluted in 30ul RNase-free water. All RNAs were checked on a NANODRP1000 and Agilent bioanalyzer 2100 RNA NANO analysis for RNA yield, purity, and integrity. The KAPA mRNA HyperPrep Kits (KK8580) with the IDT forIlluminaDual Index Adapter kit (Cat. #20021454) were used for library preparation per the manufacturer's protocol. Extra hemoglobin RNA removal and rRNA depletion using the Kapa specific reagents (RiboErase Globin) were added to the protocol. Briefly, mRNA was captured using magnetic oligo-dT beads then RiboErase Globin capture beads were applied. Fragmentation was performed using heat and magnesium. 1st strand cDNA synthesis was completed using random priming. Combined 2nd strand synthesis and A-tailing, adapter ligation, library amplification, and KAPA Pure Beads clean-ups were performed for library preparation. The strand marked with dUTP is not amplified, allowing strand-specific sequencing. All final libraries were checked on Agilent's bioanalyzer 2100 High Sensitivity DNA Chip. An equal amount of cDNA library from each sample was pooled for sequencing on the Illumina Hiseq 4000 platform.