Extended Data Fig. 3: Increased SM BCAA oxidation is specific in a fasted state and does not affect body weight or glucose tolerance on Western diet.

a, Body weights of mice; n = 8 Bckdk^l/l (WT) and n = 9 Bckdk^l/l-HSA-CreER (Cre). b, Plasma BCAA levels, c, Ratio of 3-HIB/valine, and d, Relative amount of 3-HIB in mice after a 5-hr fast (fasted) and in a fed state collected at 10 pm (fed); n = 12 WT and n = 17 Cre. e, Fasting plasma pool size of various amino acids, and f, Steady-state plasma labeling of BCAAs by U¹³C-BCAAs (fraction) during steady-state infusion of U¹³C-BCAAs before (basal, 5-hr fasted) and during (clamp, 7-hr fasted) HIEC in n = 6 WT and n = 6 Cre mice. g, Normalized labeling of TCA cycle intermediates by U¹³C-BCAAs in liver, h, iBAT, and i, gWAT at the end of the infusion and HIEC (7-hr fasted); n = 6 WT and n = 6 Cre. j, BCKDH complex activity measured in fasted quad samples fed HFD for 4 weeks; n = 4 WT and n = 3 Cre. k, 2 g/kg glucose IP-GTT in 5-hr fasted mice fed Western diet for 5 weeks; n = 8 WT and n = 9 Cre. l, Area-under-the-curve (AUC) for the GTT in k. Mice used in these experiments were male, aged 18–24 weeks and fed Western diet for 5–12 weeks. Data are presented as means ± SEM. Comparisons of two groups use two-tailed Student’s t-test with significance defined as: *p < 0.05, **p < 0.01, ***p < 0.001. Experiments with multiple comparisons at different time-points use two-way ANOVA with repeated measures. Exact p-values: b, valine fasted p < 0.000001, leucine fasted p < 0.000001, isoleucine fasted p < 0.000001, isoleucine fed p = 0.029124. c, fasted p = 0.013891. f, valine basal p = 0.030476, isoleucine basal p = 0.046680. g, glutamate p = 0.004926. h, malate p = 0.034456. I, succinate p = 0.039056.