Figure 5.

Mutation of PPARδ F269A abolishes its interaction with SMRT and the repression of PPAR isoform activity. (A) Mutation of Phe-269 disrupts PPARδ-SMRT interaction in mammalian two-hybrid assay. (B) The PPARδ F269A is not associated with SMRT in a coimmunoprecipitation assay. Two hundred ninety-three cells were transfected as indicated. The cell lysates were immunoprecipitated with anti-Flag antibody and immunoblotted with anti-HA antibody. An aliquot of the lysate input was assayed in parallel by Western blot with HA-specific antibody. (C) The PPARδ F269A does not interact with SMRT in a GST-pull-down assay. (D and E) The PPARδ F269A mutant loses its ability to repress PPARα (D) and PPARγ (E) transcriptional activity. CV-1 cells were transfected as indicated and treated with solvent, Wy (D), or 15d-PGJ2 (E) for 24 h.