Fig. 6

Effects of human preadipocyte exposure to NP before differentiation (P exposure) on mRNA and protein expression levels by real-time PCR and western blot analysis. Human preadipocytes were incubated in PAM in the presence of NP (0 or 20 μM) for 12 h. After exposure to NP, the medium was changed to differentiation medium (PADM) in the presence of PF915275 (0, 2 or 4.5 μM). a, b and c: After 6 days of incubation, PPARγ, PPARα and FASN mRNA levels in the cells were determined by real-time PCR analysis, and the values were normalized against 18 s rRNA. d, e, f: After 6 days of incubation, PPARγ (57 kDa), PPARα (57 kDa) and FASN (273 kDa) protein levels in the cells were determined by western blot analysis. The representative immunoblot was shown in the upper part, and densitometry for PPARγ, PPARα or FASN normalized to GAPDH (for PPARγ, PPARα) or Vinculin (for FASN) was shown in the bottom part. The values, which are means of independent cultures, are shown as the means ± S.E.M. and are presented as the fold-change relative to control cells (cells were not primed with NP and differentiated with PADM only). +, ++: p < 0.05, 0.01, cells with priming with NP and differentiation with PADM compared with control cells. *: p < 0.05, cells with priming with NP and differentiation with PADM containing PF compared with cells with priming with NP and differentiation with PADM only