Fig. 1: PKCα-mediated PPARγ phosphorylation at T166 can be inhibited by CDDO.

a Differential HDX mass spectrometry data for PPARγ (PDB:3e00) with rosiglitazone (RSG) and CDDO. The ribbon diagram is colored according to HDX stabilization/destabilization. Percentages of deuterium difference are color-coded according to the color gradient key. b Sequence alignment of the PKCα catalytic consensus site in PPARγ, which is conserved in mammals. c Schematic representation of the PPARγ DBD revealing that T166 is located between two zinc finger structures. d Detection of PPARγ phosphorylation at T166 in HEK293T cells with overexpressing WT, TA mutant, or TD mutant PPARγ, or without overexpression (Mock). Experiments were repeated three times. e Annotation of MS/MS spectra of the peptides of PPARγ phosphorylated at T166. f Treatment of HEK293T cells overexpressing WT PPARγ with RSG or CDDO followed by the detection of p-T166. Experiments were repeated three times. g Co-immunoprecipitation of WT PPARγ and exogenous PKCα from HEK293T cells treated with PMA (100 nM) and Ro (2 μM), RSG (1 μM), or CDDO (100 nM). Experiments were repeated three times. Data were analyzed by one-way ANOVA followed by Tukey’s test (d, f, g). Data are presented as mean ± SEM. *P < 0.05. **P < 0.01, ***P < 0.001.