Quantification of mRNA and miRNA Expression

The QIAGEN RNeasy kit was used for quantifying mRNA, as described previously (19 (link)). For complementary DNA synthesis, a Verso cDNA kit (Thermo Fisher Scientific) was used per manufacturer’s instructions. mRNA levels were measured using a 7500 Fast Real-Time polymerase chain reaction (PCR) System (Applied Biosystems) with SYBR Green-based PCR for all genes as described previously (19 (link)). The specific primers used are listed in Supplementary Table S3. Semi-quantitative real-time PCR analysis of mRNA levels was performed with reverse-transcribed RNA and 1 μM sense and antisense primers in a total volume of 20 μL. For microRNA (miRNA) quantification, total RNA was isolated from cells using TRIzol extraction reagent (Invitrogen). For mature miRNA quantification, we used TaqMan miRNA assays (Life Technologies), and real-time reverse-transcription PCR was carried out per manufacturer’s instructions. Precursor miRNAs in cells were quantified using miScript precursor miRNA assays (QIAGEN). RNU6B (for mature miRNAs) and 18S (primary and precursor miRNAs) were used as housekeeping genes.