Fig. 3: G49 induces a rapid and transient lipid overload in the liver and increases ketogenesis and FGF21.

A TGs (t0, n = 9; t1–3 h, n = 7; t6 h, n = 9; t12 h, n = 5; t24 h, n = 7; t48–72 h, n = 6; t168 h, n = 7) and Cd11b⁺Ly6g⁺ cells (n = 6) in livers. Liver TGs: 0 h vs. 3 h, ⁺⁺⁺p < 0.0001; 0 h vs. 6 h, ⁺⁺⁺p < 0.0001; 0 h vs. 12 h, ⁺⁺⁺p = 0.0004; 0 h vs. 72 h, ⁺p = 0.0118; 0 h vs. 168 h, ⁺⁺⁺p < 0.0001. One-way ANOVA with Bonferroni post hoc test. Cd11b⁺Ly6g⁺ cells, ***p < 0.0001. Unpaired two-tailed t-test. B Representative H&E (Scale bar 500 µm) and oil red O (Scale bar 50 µm) staining images (n = 4). C Liver-to-body weight ratio (t0, n = 12; t1 h, n = 6; t3 h, n = 7; t6 h, n = 15; t12–24 h, n = 6; t48–72 h, n = 5; t168 h, n = 10). 0 h vs. 6 h, ⁺⁺p = 0.0023; 0 h vs. 24 h, ⁺p = 0.0172; 0 h vs. 48 h, ⁺p = 0.0106; 0 h vs. 168 h, ⁺⁺⁺p < 0.0001. One-way ANOVA with Bonferroni post hoc test. D Plasma ALT (t0, n = 8; t6 h, n = 9; t24 h, n = 7; t168 h, n = 8). 0 h vs. 6 h, ⁺⁺⁺p = 0.0004; 0 h vs. 168 h, ⁺⁺p = 0.0016. One-way ANOVA with Bonferroni post hoc test. E mRNA levels (HFD, n = 6; t6 h, n = 6; t1 week, n = 7). Srebf1: 0 vs. 6 h, ***p < 0.0001. Fasn: 0 vs. 6 h, ***p < 0.0001. One-way ANOVA with Bonferroni post hoc test. F Plasma insulin (t0, n = 7; t6 h, n = 10, t12–168 h, n = 6). 0 h vs. 6 h, ⁺⁺⁺p < 0.0001. One-way ANOVA with Bonferroni post hoc test. G GSIS in pancreatic islets from DIO mice treated ex vivo with G49 (n = 4 islets  from independent mice). Left panel. HFD vs. H + G49: 16.7 mM, ***p < 0.0001; 2.8 vs. 16.7 mM: HFD, ⁺p = 0.0277; H + G49, ⁺⁺⁺p < 0.0001. Right panel. HFD vs. H + G49: 16.7 mM, ***p < 0.0001; 2.8 vs. 16.7 mM: HFD, ⁺⁺⁺p < 0.0001; H + G49, ⁺⁺⁺p < 0.0001. Two-way ANOVA with Bonferroni post hoc test. H Plasma TGs (t0, n = 8; t1–3 h, n = 6; t6 h, n = 9; t12 h, n = 6, t24–48 h, n = 8; t72 h, n = 6; t168 h, n = 8). 0 h vs. 24 h, ⁺⁺⁺p < 0.0001; 0 h vs. 168 h, ⁺⁺⁺p = 0.0002. One-way ANOVA with Bonferroni post hoc test. I RER during 3 days of treatment (n = 4 HFD; H + G49). Light: HFD vs. H + G49: ***p < 0.0001 at day 3; H + G49: day 0 vs. day 3, ⁺⁺p = 0.0059. Dark: HFD vs. H + G49: *p = 0.0246 at day 1, **p = 0.0042 at day 3. RER analyzed 6 h after first and second G49 injection (n = 4). HFD vs. H + G49: **p = 0.0066 after 6 h. Two-way ANOVA with Bonferroni post hoc test. J Cpt1a (t0, n = 6; t12–24 h; n = 8; t48–168 h, n = 7). 0 h vs. 24 h, ⁺p = 0.0247; 0 h vs. 72 h, ⁺⁺⁺p = 0.0001; 0 h vs. 168 h, ⁺⁺⁺p < 0.0001. Ppara (t0, n = 7; t6 h, n = 9; t12 h, n = 6; t24 h, n = 5; t48 h, n = 6; t72 h, n = 5; t168 h, n = 6). 0 h vs. 48 h, ⁺⁺p = 0.0015; 0 h vs. 72 h, ⁺⁺⁺p < 0.0001; 0 h vs. 168 h, ⁺⁺⁺p < 0.0001. One-way ANOVA with Bonferroni post hoc test. K Fatty acid oxidation (FAO) in livers at 72 h (HFD, n = 7; H + G49, n = 8). ASM, **p = 0.0068; CO₂, **p = 0.0037. Unpaired two-tailed t-test. L Hmcs2 (t0, n = 6; t6 h, n = 9; t12–24 h, n = 6; t48–168 h, n = 7). 0 h vs. 12 h, ⁺⁺p = 0.0045; 0 h vs. 24 h, ⁺⁺⁺p = 0.0001; 0 h vs. 48 h, ⁺⁺p = 0.0063; 0 h vs. 72 h, ⁺⁺⁺p < 0.0001; 0 h vs. 168 h, ⁺⁺⁺p = 0.0002. Blood ketone bodies (HFD t0, n = 15; t6 h, n = 11; t12–72 h, n = 10; t168 h, n = 9 and H + G49 t0, n = 10; t6 h, n = 12; t12–72 h, n = 10; t168 h, n = 8). HFD vs. H + G49: 12 h, ***p < 0.0001; 24 h, ***p < 0.0001; 48 h, **p = 0.0026; 72, 168 h, ***p < 0.0001. Two-way ANOVA with Bonferroni post hoc test. M Fgf21 (t0, n = 5; t12–168, n = 7). 0 h vs. 12 h, ⁺p = 0.0432; 0 h vs. 24 h, ⁺⁺⁺p < 0.0001; 0 h vs. 48 h, ⁺p = 0.0289; 0 h vs. 72 h, ⁺⁺p = 0.0070; 0 h vs. 168 h, ⁺p = 0.0255. Plasma FGF21 (t0, n = 6; t12–168 h; n = 7). 0 h vs. 12 h, ⁺⁺p = 0.0078; 0 h vs. 24 h, ⁺⁺p < 0.0064; 0 h vs. 48 h, ⁺⁺⁺p = 0.0002; 0 h vs. 72 h, ⁺p = 0.0101; 0 h vs. 168 h, ⁺p = 0.0258. Brown–Forsythe and Welch ANOVA with Dunnett’s T3 post hoc test. Data are mean ± SEM. Source data are provided as a Source Data file.