Quantitative RT-PCR analysis

Extraction of RNA was performed using Trizol (Invitrogen) according to the manufacturer’s instructions. Total cDNA was synthesized from isolated RNA using M-MLV reverse transcriptase (Takara Bio Company, Otsu, Japan). Samples were examined by quantitative PCR using a QuantiTect SYBR Green PCR Kit (Qiagen, CA, USA) and the following primers: glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 5′-GGC TGC TTT TAA CTC TGG TA-3′ and 5′-ACT TGA TTT TGG AGG GAT CT-3′), mouse PPARγ (5′-CCA TTC TGG CCC ACC AAC-3′ and 5′-AAT GCG AGT GGT CTT CCA TCA-3′), human PPARγ (5′-TTC AGA AAT GCC TTG CAG TG-3′ and 5′-CCA ACA GCT TCT CCT TCT CG-3′), aP2 (5′-CAC CGC AGA CGA CAG GAA G-3′ and 5′-GCA CCT GCA CCA GGG C-3′), 36B4 (5′-AGA TGC AGC AGA TCC GCA T-3′ and 5′-GTT CTT GCC CAT CAG CAC C-3′), and C/EBP-α (5′-GCG GGC AAA GCC AAG AA-3′ and 5′-GCG TTC CCG CCG TAC C-3′). The expression level of each gene was determined from 3T3-L1 and MEF cell samples and was normalized to the expression of the 36B4 gene in the same sample. The expression levels of genes in PC-3 cells were normalized to those of GAPDH.