GSM5903310: WT_IL25_RNA rep1; Homo sapiens; RNA-Seq
Total RNA was extracted from HSMCs that were untreated, treated with various stimuli using the RNeasy mini kit (Qiagen, Valencia, CA) according to the manufacture's protocol. The quality of input RNA was analyzed by running the RNA sample on 4150 TapeStation System (Agilent, CA) PolyA mRNA was isolated using the NEBNext® Poly(A) mRNA magnetic isolation module (E7490, New England Biolabs, Ipswich, MA). RNA-seq libraries were obtained using the NEB-Next® UltraTM II RNA library prep kit for Illumina® (E7770S, New England Biolabs, Ipswich, MA). The isolated mRNA was fragmented, reverse transcribed into cDNA, end-repaired, and adapter-ligated. The ligated samples were digested by USER enzyme, amplified by PCR and purified using AMPure-XP beads (Beckman Coulter Life Sciences, A63881, Indianapolis, IN). The quantity and quality of RNA libraries were assessed on an Agilent Technologies 2100 Bioanalyzer.