Real-Time qRT-PCR Analysis of miRNA and mRNA

Real-time qRT-PCR analysis was performed as previously described11 (link)45 (link). Briefly, RNA was extracted using miRNeasy columns (Qiagen, Valencia, CA). miRNA expression analysis was performed using the qScript miRNA cDNA synthesis kit (Quanta Biosciences, Gaithersburg, MD) and PerfeCTa SYBR Green Supermix (Quanta Biosciences). miRNAs were amplified using specific mature miRNA sequences as forward primers and the universal primer provided in the kit as the reverse primer. U6 was used as internal control. A GeneAmp RNA PCR kit (Applied Biosystems, Carlsbad, CA) and POWER SYBR Green mix (Applied Biosystems) were used for mRNA quantification. PCR primer sequences are in Supplementary Table 3.