RNA was purified from frozen aortic tissues using the RNeasy Fibrous Tissue Kit (Qiagen). RNA amounts were quantitated by Qubit fluorometer (Life Technologies) and RNA quality was assessed using an Agilent 2100 Bioanalyzer. Total RNA preparations with ribosomal integrity numbers >8 were used for RNA-Seq experiments performed using Illumina HiSeq4000. Libraries were prepared from 1μg of total RNA using TruSeq Stranded mRNA kit protocol (Illumina®) according to the supplier's recommendation. Briefly, poly-A RNA was isolated using oligo-dTs attached to magnetic beads, fragmented into ~300 bp-long pieces using divalent cations under elevated temperature, used as template for double-stranded cDNA synthesis, Illumina adapters ligation and PCR-mediated cDNA library amplification. Sequencing of random paired-end 75 bp molecules was carried out on Illumina HiSeq4000.
GEO Accession
GSM2949418