Quantification of mRNA and miRNA Expression

Total RNA was extracted from cells using RNA isolater (Vazyme, Nanjing, China). Complementary DNA (cDNA) for mRNA or miRNA was then synthesized with HiScript RT SuperMix (Cat # R223-01, Vazyme). MiRNA/U6 snRNA (small nuclear RNA) RT primer mix and PCR specific primer set (GenePharma, China) were used to reversely synthesize cDNA for miRNAs, detect and quantify miRNAs expression, respectively. qRT-PCR was performed with qPCR SYBR® Green Master Mix (Vazyme) on Real-Time PCR system (Applied Biosystems, USA). U6 snRNA or GAPDH was used as an endogenous control for miRNA or mRNA, respectively. 2^−∆∆Ct method was carried out to measure the relative expression levels of mRNA or miRNA. The primer sequences were mentioned in our previous study [23 (link)].