RNA was extracted using RNeasy Lipid Tissue Mini Kit (QIAGEN, Germantown, Maryland) and processed as done previously (23 (link)). Samples were pooled into single library (n = 8 pooled samples per library) and sequenced using the NextSeq 500 High Output Kit (300 cycles, paired end 100bp) on the Illumina NextSeq 500 platform. Sequenced reads were assessed for quality using the Illumina Basespace Cloud Computing Platform and FASTQ sequence files were used to align reads to the rat reference genome [Rattus norvegicus/Rn5 (Refseq)] using RNA-Seq Alignment Application with STAR aligner. Fragments per kilobase of transcript per million mapped reads (FPKM) values of reference genes and transcripts were generated using Cufflinks 2. Differential expression was characterized by a fold change threshold of ≥1.5 as previously done (24 (link)). Relative expression of selected genes was determined using predesigned TaqMan gene expression assays as described in the online supporting information.