Figure 5.

CAB inhibited the recruitment of PPARγ coactivator. (A) Dose-dependent effects of CAB on the ability of PPARγ coactivator recruitment as evaluated by UAS/Gal4 system. HEK293T cells were co-transfected with Gal4-PPARγ-LBD and the UASE1b-TATA-Luc reporter gene, while the control plasmids were treated with rosiglitazone (0.5 μmol/L) and increasing concentrations of CAB for 18 h. The cells were harvested for luciferase and β-galactosidase assays. The values shown are the means±standard deviations from three independent experiments. (B) Dose-dependent effects of CAB on the recruitment of PPARγ transcription co-activator CBP as determined by yeast two-hybrid system. The overnight cultures of yeast cells containing pGADT7-CBP and pGBKT7-PPARγ were diluted with fresh media to an initial OD₆₀₀ of 0.3 and treated with rosiglitazone (10 μmol/L) and increasing concentrations of CAB or DMSO (as vehicle control) for 16 h at 30 °C. Relative α-galactosidase activity was determined by OD₄₁₀/OD₆₀₀. The values shown are the means±standard deviations from three independent experiments.