Abi quantstudio 5 instrument

RNA iso (TaKaRa, Otsu, Japan) was used to extract total RNA from muscles and cells according to the instructions. Reverse transcription to synthesize RNA cDNA was performed using HiScript Q-RT SuperMix for qPCR (+gDNA wiper) (Vazyme, Guangzhou, China). Using the ABI QuantStudio 5 instrument (Thermo Fisher Scientific, Fremont, NY, USA), the ChamQ Universal SYBR qPCR Master Mix (Vazyme) was used to perform qRT-PCR to detect the expression level of mRNAs. Data analysis of the results used the 2^−∆∆Ct method to calculate the relative expression of genes. The primers for real-time qPCR were designed by Premier Primer 5.0 software (Premier Biosoft, Palo Alto, CA, USA). Primers used for qPCR are listed in Table S5.

The MagZol Reagent (Magen, Guangdong, China) was used to extract total RNA following the manufacturer’s protocol. The total RNA was used to synthesize cDNA by HiScript II Q RT SuperMix for qPCR (+gDNA wiper) (Vazyme, Nanjing, China) according to the manufacturer’s protocol. Quantitative PCR (qPCR) was performed using ChamQ Universal SYBR qPCR Master Mix (Vazyme, Nanjing, China) on ABI QuantStudio 5 instrument (Thermo Fisher, Grand Island, NY, USA). The primers used in qRT-PCR were designed by Primer Premier 6.0 software and synthesized by TSINGKE (Beijing, China). The primers are listed in Table S1.