Extended Data Figure 3. Increased BCAA oxidation in BCKDK muscle knockout 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 10pm (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, 2g/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. 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.