Fig. 2: A single dose of G49 rapidly induces a beiging immune signature in eWAT from DIO mice.

A Representative H&E staining images and morphometric analysis of adipocyte area (n = 8 mice in HFD and H + G49). Scale bar 100 µm. Representative TUNEL images and quantification (HFD, n = 5; H + G49, n = 8). Scale bar 50 µm. ***p < 0.0001. Unpaired two-tailed t-test. B Fgf21 mRNA (HFD, n = 5; H + G49, n = 6) **p = 0.0015. Unpaired two-tailed t-test. mRNA levels of remodeling-related genes (Acta2, HFD, n = 8; 6 h, n = 6; 1 week, n = 8; Tgfb, HFD, n = 9; 6 h, n = 6; 1 week, n = 7; Mmp9, HFD, n = 8; 6 h, n = 7; 1 week, n = 6) in eWAT from DIO mice treated with G49 for 6 h or 1 week. Acta2: 0 h vs. 6 h, **p = 0.0023. Tgfb: 0 h vs. 6 h, **p = 0.0012. Mmp9: 0 h vs. 6 h, ***p = 0.0001. One-way ANOVA with Bonferroni post hoc test. C Representative F4/80 and TH immunostaining images in eWAT (n = 4). Scale bar 100 µm. D Gene expression-related beiging signature analyzed 6 h after G49 injection. Left panel eWAT: Arg1, HFD, n = 8; H + G49, n = 7, ***p < 0.0001; Il10, HFD, n = 7; H + G49, n = 8, ***p = 0.0003; Tgfb, HFD, n = 8; H + G49, n = 8, **p = 0.0021; Th, HFD, n = 6; H + G49, n = 7, **p = 0.0012. Unpaired two-tailed t-test. Middle panel SVF: Th, HFD, n = 6; H + G49, n = 7, *p = 0.0149. Unpaired two-tailed t-test. Right panel SVF gene expression at different time-points Il4, 0 h, n = 6; 6 h, n = 7; 24 h, n = 5; 72 h, n = 6. Il10, 0 h, n = 6; 6 h, n = 6; 24 h, n = 5; 72 h, n = 6. Il13, 0 h, n = 5; 6 h, n = 7; 24 h, n = 5; 72 h, n = 7. Il4: 0 h vs. 6 h, **p = 0.0015. Il10: 0 h vs. 6 h, ***p = 0.0007. Il13: 0 h vs. 6 h, **p = 0.0017. One-way ANOVA with Bonferroni post hoc test. E–I Immune cell populations in SVF from eWAT at 6 h post-G49 injection (n = 6 in HFD; H + G49). SiglecF⁺Cd11b⁺Cd11c⁻ (**p = 0.0099) and IL4⁺ cells (***p < 0.0001) (E), Cd11b⁺F4/80⁺Cd206⁺ and Cd11b⁺F4/80⁺Cd11c⁺ cells and ratio between these populations (**p = 0.0036) (F), Cd11b⁺Ly6g⁺ cells (*p = 0.0331) (G), Cd11b⁺Ly6g⁺Cd206⁺ and Cd11b⁺Ly6g⁺Cd11c⁺ cells and ratio between these populations (**p = 0.0031) (H), and TCRβ⁺NK1.1⁺Cd3⁺ cells (**p = 0.0024) (I). E–I Unpaired two-tailed t-test. Data are mean ± SEM. Source data are provided as a Source Data file.