Quantitative real-time RT-PCR analysis
Total RNA was extracted from liver, muscle, hypothalamus, and periepididymal fat tissues using Trizol reagent (Invitrogen, Carlsbad, CA, USA) and purified with RNeasy Mini Kit (Qiagen, Hilden, Germany) following manufacturer’s protocol. cDNA was synthesized using ReverTra Ace qPCR RT Master Mix (Toyobo, Osaka, Japan). A total of 68 genes were chosen; if primer information was not available in the literature, primer was designed using PRIMER3 software. GAPDH was used for normalization. The primer sequences are provided in supplementary Table 1. qRT-PCR was performed using QuantiSpeed SYBR Green Kit (PhileKorea, Republic of Korea).