Figure 7
Madecassic acid as a PPARγ agonist. (a and b) EL-4 cells were treated with madecassic acid (1, 3, 10 μM) and rosiglitazone (2 μM) for 24 h. The expression of the PPARγ-responsive genes CD36 and LPL was quantified by western blot and real-time PCR. (c and d) EL-4 cells were treated with madecassic acid (3 μM), GW9662 (1 μM) and T0070907 (1 μM) for 24 h. The expression of CD36 and LPL was measured by western blot and real-time PCR. (e and f) EL-4 cells were treated with madecassic acid (3 μM) and PPARγ siRNA for 24 h. The expression levels of CD36 and LPL were measured by western blot and real-time PCR. (g) The docking between madecassic acid and PPARγ-LBD in EL-4 cells. (h) Binding of madecassic acid to PPARγ-LBD in a competitive TR-FRET assay. (i) The effect of madecassic acid on the nuclear accumulation of PPARγ in EL-4 cells was determined by an EMSA in EL-4 cells. (j) The effect of madecassic acid on PPARγ-dependent transactivation in EL-4 cells was assayed with a luciferase reporter gene assay. GAPDH was used as a cytoplasmic marker. The data were expressed as means±S.E.M., n=3. *P<0.05, **P<0.01 versus control group, $P<0.05, $$P<0.01 versus madecassic acid group
