Fig. 8 |. Double knockout of BCKDK in both SM and liver has no effect on insulin sensitivity.

a, Western blotting of fasted tissues from n = 3 WT and n = 3 BCKDK double knockout mice. Liver BCKDK is the bottom band, as shown by the black arrow. The top band is non-specific; 14-3-3 is the loading control. b, IP-GTT (1.5 g kg⁻¹) in mice fasted for 5 h and fed HFD for 6 weeks; n = 10 WT and n = 10 double knockout mice. c, AUC for the GTT in b; n = 10 WT and n = 10 double knockout mice. d, Insulin concentration at t = 0 and t = 20 min (n = 10 WT and n = 10 double knockout mice) during the GTT from b. e, Plasma BCAA concentration in HFD-fed mice fasted for 5 h or refed for 2 h after overnight fasting; n = 10 WT, n = 6 Bckdk^loxP/loxP-HSA-CreER (SM knockout) and n = 10 double knockout mice. f, Western blotting of refed tissues from n = 3 WT and n = 3 double knockout mice; 14-3-3 is the loading control. g, IP-GTT (1.5 g kg⁻¹) in mice fasted for 5 h and fed HFD for 7 weeks, treated with 100 mg kg⁻¹ of either vehicle or BT2 the night before and the morning of the GTT; n = 8 BCKDK double knockout + vehicle and n = 8 BCKDK double knockout + BT2 mice. h, AUC for the GTT in g. i, Insulin concentration at t = 0 and t = 20 min (n = 8 BCKDK double knockout + vehicle and n = 8 BCKDK double knockout + BT2 mice) during the GTT from g. j, Plasma BCAA concentration in mice fasted for 5 h or refed for 2 h after overnight fasting, treated with 100 mg kg⁻¹ of either vehicle or BT2 the night before and the morning of plasma collection; n = 8 BCKDK double knockout + vehicle and n = 9 BCKDK double knockout + BT2 mice. Mice used for these experiments were 13–23-weeks old and were fed HFD for 6–7 weeks. Data are presented as the mean ± s.e.m. When two groups were compared, a two-tailed Student’s t-test was used, with significance defined as *P < 0.05 and **P < 0.01. In the experiments with multiple comparisons at different time points, a repeated measures, two-way ANOVA was used. e, Valine WT versus double knockout fasted P = 0.047907; valine WT versus SM knockout fasted P = 0.049487; leucine WT versus double knockout fasted P = 0.020429; leucine WT versus SM knockout fasted P = 0.031846; isoleucine WT versus double knockout fasted P = 0.014609; isoleucine WT versus SM knockout fasted P = 0.031719. h, P = 0.039819. j, Valine fasted P = 0.025933, valine refed P = 0.003017; leucine fasted P = 0.022498, leucine refed P = 0.004677; isoleucine fasted P = 0.041485, isoleucine refed P = 0.003193.