Quantification of miRNA and mRNA Expression

Total RNA was extracted as previously defined. For mature miRNAs, All-in-One™ miRNA First-Strand cDNA Synthesis Kit (Genecopoeia) was used to generate 10 μL cDNA. Then using miR-specific primers and universal adaptor PCR primers (Genecopoeia), qRT-PCR was performed with All-in-One™ miRNA qRT-PCR Detection Kit (Genecopoeia) on CFX Connect Real-Time System (Bio-Rad). For mRNA, the reverse transcription was carried out using HiScript II Q RT SuperMix (Vazyme), and qRT-PCR was performed with ChamQ™ SYBR Color qPCR Master Mix (Vazyme) on CFX Connect Real-Time System (Bio-Rad) [21 (link)]. Specific primers for qRT-PCR of mRNA were listed in Supplementary Data. The U6 small nuclear RNA or GAPDH was used as an endogenous control. Each experiment was repeated at least three times and the results were analyzed using 2^-△△CT method.