RNA purification and real-time reverse transcription polymerase chain reaction (qRT-PCR)

Total RNA was obtained from cultured cells with TRIzol reagent (Invitrogen, USA) as instructed by the manufacturer. Quantitative real-time RT-PCR detecting mature miR-30e was carried out in triplicate with RT Reagent Kit (Vazyme, Nanjing, China) as directed by the manufacturer, withAceQ SYBR Master Mix (Vazyme, Nanjing, China) on a 7900HT system. MicroRNA-30e levels in each group were determined relative to U6 amounts, by the 2^−ΔΔCt) method.