Figure 2.

The FPLD-associated L451P mutant, but not the colon cancer-associated mutant K450Q, displays impaired transcriptional activity. A-D. U2OS cells were transiently cotransfected with expression vectors encoding PPARγ2 WT or mutants (A, left panel and C), PPARγ1 WT or mutants (A, right panel) or PPARα WT or S414P mutant (C). The activation of 3× peroxisome proliferator response element (PPRE)-Tk-Luc reporter (A and C), Cidec- (B), or Fabp4-Luc reporter (B), in the absence or presence of 1 μM rosiglitazone (PPARγ) or 100 μM Wy14643, is expressed as fold induction over that with empty vector (EV). D. U2OS cells were transfected with equal amounts of plasmids harboring PPARγ2 WT, L451P, and L496A/E499A in absence and presence of 1 μM rosiglitazone. Results are averages of at least three independent experiments assayed in duplicate ± SEM. *P < 0.05; **P < 0.01 cells transfected with mutant vs. WT. Expression levels of the different proteins were confirmed by western blot analysis using a PPARγ or PPARα specific antibody. The arrow indicates PPARγ and the asterisk indicates an unknown non-specific band. WT, wildtype.