2.4. RNA Isolation and qRT-PCR Analysis

Total RNA was isolated using TRIzol reagent (Thermo Fisher Scientific, USA) according to the instructions provided. First-strand cDNA was generated with a PrimeScript RT Reagent Kit with gDNA Eraser (TaKaRa, Japan), and miRNA reverse transcription was performed using a Mir-X miRNA qRT-PCR SYBR Kit (Clontech, Japan). Real-time PCR was performed using the PrimeScript RT Reagent Kit and SYBR Premix Ex Taq (TaKaRa, Japan) on a CFX96 Real-Time System (Bio-Rad, USA) with the reaction conditions provided in the instructions. The primer details used in the study were included in Table S2. Additionally, the common miRNA mRQ 3′ primer was provided in the Mir-X miRNA qRT-PCR SYBR Kit (Clontech, Japan).