Figure 6.

BCKDH deficiency impacts differentiation and glycolytic flux in human adipocytes. Characterization of control (WT) and BCKDHA knock-out (KO) human adipocytes. A, Western blot of BCKDHA (three cellular replicates). B, gene expression of BCKDHA (12 cellular replicates) C, gene expression of differentiation markers (9-12 cellular replicates). D, bodipy staining of lipid droplets. Scale bars, 100 μm and 25 μm (inlay). E, Glucose uptake (13 cellular replicates). F and G Extracellular lactate (14 cellular replicates) and metabolite levels (13 cellular replicates) in media following 24 h of culture. H, intracellular metabolite levels 24 h post media change (14 cellular replicates). Data are presented as means ± SD. For A and D, results are depicted from one representative experiment which was repeated independently at least two-three times. For (B–C) and (E–H), all cellular replicates were collected over two-three separate differentiation rounds. Two-tailed students t test was used to test significant differences in (B–C) and (E–H). Multiple unpaired t-tests with correction for multiple comparison using the Holm-Sidak method was used in (G–H). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.