Extended Data Fig. 7: Liver BCKDK knockout does not affect body weight or glucose tolerance at various time points on HFD or after a BCAA challenge.

a-c, Western quantification of BCKDK in tissues normalized to loading control; n = 3 Bckdk^l/l-AAV8:GFP (GFP) and n = 3 Bckdk^l/l-AAV8:Cre (Cre). d-f, Western quantification of the ratio pBCKDH/BCKDH; n = 3 GFP and n = 3 Cre. g, Body weights of n = 9 GFP and n = 10 Cre mice on HFD. h, Plasma labeling of BCAAs after U¹³C-BCAAs gavage (fraction) 5-hr fasted mice; n = 4 GFP and n = 5 Cre. Labeling fraction of TCA cycle intermediates 10 min post U¹³C-BCAA gavage in i, quad j, iBAT, & k, gWAT in 5-hr fasted mice; n = 4 GFP and n = 5 Cre. l, BCKDH complex activity assay measured in fasted livers from n = 4 GFP and n = 3 Cre mice. m, 1.5 g/kg glucose IP-GTT in 5-hr fasted mice fed HFD after a BCAA gavage given at t = 0; n = 8 GFP and n = 10 Cre. n, Area-under-the-curve (AUC) for the GTT in m. o, Plasma BCAA concentration of 5-hr fasted plasma before (t = 0) (n = 8 GFP and n = 9 Cre) and 20 min after (t = 20) (n = 8 GFP and n = 10 Cre) a BCAA gavage from GTT + BCAA gavage in m. Mice used in these experiments were male, aged 8–26 weeks and on HFD for 2–12 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 and **p < 0.01. Experiments with multiple comparisons at different time-points use two-way ANOVA with repeated measures. Exact p-values: a, p = 0.029151. d, p = 0.005367.