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. 2010 Oct;177(4):1697–1709. doi: 10.2353/ajpath.2010.090789

Figure 4.

Figure 4

Involvement of PPARα activation in adiponectin (APN)-mediated heme oxygenase (HO)-1 induction. APN increased PPRE enhancer-driven luciferase activity (A) and nuclear translocation of PPARα activation (B) and subsequent HO-1 induction (C). A: HEK 293 cells were transiently simultaneously transfected with pBV-Luc-PPARα enhancer and Renilla control vectors or with additional transfection of pcDNA-PPARα, and pcDNA-RXR (0.5 μg/well) for five hours, after which fresh medium was added for overnight inoculation. Cell lysates were harvested after cells were treated with APN for six hours. B: Cells were respectively pretreated with either AMPK or PPARα antagonist, or a PPARα agonist for one hour before the administration of APN for another one hour for nuclear translocation of PPARα by Western blot analysis and (C) subsequent HO-1 induction at six hours of APN treatment. D: Elimination by PPARα knockdown of APN-mediated HO-1 induction. The method of cells with PPARα knockdown was described in Materials and Methods. Equal loading or transfer was confirmed by incubation with an anti-GAPDH or anti-lamin A/C antibody. Representative results of three separate experiments are shown, and data are presented as the mean ± SEM (*P < 0.05, **P < 0.01, and ***P < 0.001 vs. the control; †P < 0.05 vs. APN alone).