Fig. 6

VAV1-AKT-FOXO1-PPAR-γ pathway was involved in the regulation of CD226KO on macrophage polarization. A Immunoblot analysis of the expression of PPAR-γ in the epididymal fat of WT and CD226KO mice fed with either chow or a HFD for 16 weeks. Blots are representative of three independent experiments. B mRNA levels of PPAR-γ in epididymal SVF from WT and CD226KO mice on a HFD (n = 3). C Immunoblot analysis of the expression of p-VAV1, VAV1, p-AKT, AKT, p-FOXO1, FOXO1, and PPAR-γ in peritoneal macrophages from WT or CD226KO mice. Blots are representative of three independent experiments. D Luciferase activities were measured in RAW264.7 cells transfected with pGL3-PPAR-γ or pGL3-basic plasmids following 10E5 or isotype antibody treatment (n = 3). E mRNA levels of M1/M2-type cytokines and chemokines in LPS-stimulated CD226KO peritoneal macrophages treated with the PPAR-γ specific inhibitor GW9662 or DMSO vehicle control (n = 3). F Culture supernatant concentrations of proinflammatory cytokines in LPS-stimulated CD226KO peritoneal macrophages treated with GW9662. Combined data from three independent experiments (n = 3). G FACS analysis of CD11c⁺ M1 in LPS-stimulated CD226KO peritoneal macrophages treated with GW9662 (n = 3). Data represent mean ± SEM. Differences between groups were determined by one way ANOVA with Tukey’s multiple test (D) or unpaired Student’s t-test (B, E, F, G). *P < 0.05, **P < 0.01