Quantitative RT-PCR for mRNA and miRNA Analysis

Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed as previously described (Xia et al., 2019 (link)). The messenger RNA (mRNA) and miRNA were isolated from cells using RNeasy mini kit (Qiagen) according to the manufacturer’s instructions. Genomic DNA was removed using DNase I digestion kit (Qiagen). cDNA was synthesized using miScript II RT kit (Qiagen). Transcripts were amplified using specific primer sets (Supplementary Table 1) and SYBR green PCR kit (Qiagen) with the ABI7500 (Applied Biosystems). Reactions were run in triplicates for each sample and no-template blanks were used as negative controls. Values were normalized to the Gapdh (for mRNA) and U6 snRNA (for miRNA).