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Total RNA was extracted from mature 3T3-L1 adipocyte cells with TRIzol reagent (Invitrogen; Thermo Fisher Scientific, Inc.) in accordance with the manufacturer's recommended protocols. RNA samples with OD₂₆₀/OD₂₈₀ ratios >2.0 were employed for semi-quantitative RT-PCR. One microgram of total RNA was employed for the production of complementary DNA using a Maxime RT premix kit (Intron Biotechnology, Inc., Seongnam, Korea) at 45°C for 60 min and an RT-PCR system (c1000 thermal cycler; Bio-Rad Laboratories, Inc., Hercules, CA, USA). PCR amplification was performed with primers, cDNA and Taq MasterMix (MG Taq MasterMix; Doctor Protein; MGMED, Inc., Seoul, Korea), and it was performed at 95°C for 5 min, followed by 30 cycles of 95°C for 30 sec, annealing for 30 sec at 54°C and an extension step of 1 min at 72°C, and a final extension step of 5 min at 72°C. All primers used in this study are listed in Table I. The PCR products were then run on 1.5% (v/v) agarose gels and stained with ethidium bromide, and images were captured with a gel documentation and analysis system (care stream MISE; Carestream Health, Inc., Rochester, NY, USA). The expression levels were quantified using ImageJ software 1.48 (National Institutes of Health, Bethesda, MD, USA).