Figure 4: GW7845 is a selective PPARγ agonist. (A), PPAR transcriptional activity was measured by mammalian cell transfection assays. HeLa cells were transfected with a PPARγ or PPARδ expression vector in combination with a DR1-Luc reporter plasmid and a β-galactosidase normalization vector. After transfection, cells were treated with vehicle (Veh) or 1 μM carbaprostacyclin (Carb), or 1 μM TZDs: rosiglitazone (Rosi), pioglitazone (Pio), troglitazone (Trog) or ciglitazone (Cig), or 100 nM GW7845 in the absence or presence of 1 μM PPARδ antagonist GSK660 for 40 h. Cells were harvested and assayed for luciferase activity; all luciferase assay values were normalized to β-gal controls. Data points are the average of triplicate determinations in a representative experiment, and the average coefficient of variance for each value is <10%. (B) Schematic of mammalian two-hybrid assay. (C) HeLa cells were transfected with VP16 and pM control vectors, pM-ASC-2 (NR-box), and VP16-PPARγ (VP16-PPARδ or VP16-PPARγ expression vectors in combination with a 5x-Gal4- TATA-Luc reporter plasmid and a β-galactosidase normalization vector. After transfection, cells were treated, assayed and analyzed as in A.