Total RNA was extracted from fat tissues or hypothalamus using RNeasy Lipid Tissue Kit (Qiagen), and cDNA was synthesized from 2 μg of RNA using M-MLV reverse transcriptase (Invitrogen) with oligo(dT). Gene expression assay was conducted using SYBR Green methods on Viia7 (Applied Biosystems), and relative Ct values were normalized by β-actin. Primer sequences are available upon request.
