Quantifying Gene Expression in Adipocytes

RT-qPCR was measured as previously described [19 (link)]. Total RNA from the 3T3-L1 adipocytes was extracted using TRIzol reagent (GeneAll Biotechnology, Seoul, Korea), and the cDNA was synthesized from RNA extracted using M-MLV reverse transcriptase (Bioneer, Daejeon, Korea). Afterward, RT-qPCR was performed in a fluorescent thermal cycler (Rotor-Gene^TM 3000, Corbett Research, Mortlake, NSW, Australia) using Universal SYBR^® Green PCR Master Mix (Bioneer Co., Daejeon, Korea). The 2^−ΔΔCt method was used as a relative quantification strategy, and target gene expression was normalized using β-actin as an endogenous control. Primers used for RT-qPCR analysis are shown in Table 1.