Fig. 3: APC-derived EVs suppress WAT inflammaging to prevent obesity.

a A schematic diagram of the mouse treatments. b Body weight gain of 12-month-old male mice fed a NCD or HFD and treated with PBS, yEVs or mEVs (n = 10 per group). c Energy expenditure of 12-month-old male mice treated with mEVs or yEVs (n = 4 per group). GTT (d) and ITT (e) of 12-month-old male mice fed a NCD or HFD and treated with PBS, yEVs or mEVs (n = 10 per group). f Western blot of p-AKT^Ser473/AKT in the eWAT, liver and skeletal muscle of 12-month-old male mice fed a NCD or HFD and treated with PBS, yEVs or mEVs. g Confocal images and quantitative analysis of crown-like structures (CLSs, arrows) in eWAT from 12-month-old male mice fed a NCD or HFD and treated with PBS, yEVs or mEVs (n = 10 biological replicates per group). Scale bar, 25 µm. h Flow cytometry analysis of M1 (CD11b⁺/F4/80⁺/CD11c⁺/CD206^-) and M2 (CD11b⁺/F4/80⁺/CD11c^-/CD206⁺) macrophage subsets in the eWAT of 12-month-old male mice fed a NCD or HFD and treated with PBS, yEVs or mEVs (n = 10 per group). i RT‒qPCR analysis of Tnf-α, Il-1β, and Il-6 in eWAT from 12-month-old male mice fed a NCD or HFD and treated with PBS, yEVs or mEVs (n = 10 biological replicates per group). j Western blot analysis of p19^ARF, p53 in eWAT from 12-month-old male mice fed a NCD or HFD and treated with PBS, yEVs or mEVs (n = 4 biological replicates per group). k RT‒qPCR analysis of p16^Ink4a, p19^Arf, and p21 in eWAT from 12-month-old male mice fed a NCD or HFD and treated with PBS, yEVs or mEVs (n = 10 biological replicates per group). l Representative SA-β-Gal staining of the iWAT and eWAT of 12-month-old male mice fed a NCD or HFD and treated with PBS, yEVs or mEVs. The data were presented as mean ± SEM. Statistical significance was assessed by two-way ANOVA and Bonferroni’s multiple comparisons test (b, left panels in d and e), two-sided ANCOVA with body weight as co-variate (left panel in c), two-sided Student’s t-test (right panel in c), or one-way ANOVA (i, k and right panels in d, e, g, h and j). See also Supplementary Fig. 4 and Supplementary Fig. 5. Source data are provided as a Source Data file.