Chiglitazar Preferentially Regulates Gene Expression via Configuration-Restricted Binding and Phosphorylation Inhibition of PPARγ

Figure 4

Chiglitazar inhibited TNFα-induced and CDK5-mediated phosphorylation of PPARγ. (a) HPA-v cells were cultured in 10 cm plates and incubated with each agonist at the indicated concentration in the presence or absence of TNFα (5 ng/ml). Cells were lysed and then subjected to immunoprecipitation (IP) with primary anti-PPARγ antibody followed by SDS-PAGE and immunoblot analysis as described in Materials and Methods. (b) In vitro phosphorylation was performed using purified active CDK5/p25 (5 ng/mL) and purified recombinant human PPARγ-LBD. Chi, Ros, and Pio were added to the reaction, respectively, at the indicated concentrations in presence of TNFα (5 ng/ml) and then incubated for 30 minutes at 30°C. Immunoblot analysis was performed to determine the levels of the specified proteins.

(a)

(b)