Figure 6. Inhibition of BCAA catabolism impairs adipocyte differentiation.
(a) Western blot of Bckdha and β–Actin in differentiated 3T3–L1 adipocytes.
(b) Basal respiration in Control and Bckdha KD adipocytes normalized to nuclear quantitation.
(c) BCAA uptake in Control KD and Bckdha KD adipocytes.
(d) Absolute lipogenic flux of leucine to palmitate synthesis.
(e) Relative abundance of the most abundant fatty acids.
(f) Quantitative PCR analysis of adipocyte–specific gene expression. Data represents 2 biological replicates analyzed via 2–way ANOVA with Holm–Sidak’s multiple comparisons test.
(g) Representative images of adipocyte differentiation (scale bar: 200 μm) From left: Control KD, Bckdha KD1, Bckdha KD2.
Data presented in (b)–(e) represent mean ± s.d. (f) is presented as mean ± s.e.m. Asterisks in (b)–(e) represent significance compared to Control KD where *represents p<0.05, **represents p<0.01, and ***represents p<0.001. Unless otherwise specified, data is 3 technical replicates representative of 3 biological replicates analyzed via two–way ANOVA with Holm–Sidak’s multiple comparison test.