Figure 3.

Effect of DGAT inhibition on incorporation of glycerol, acetate and glucose into TAG and total lipids. Human myotubes were grown and differentiated on 12-well tissue culture plates for 6–7 days. At day 7 of differentiation myotubes were incubated with D-[¹⁴C(U)]glycerol (1 µCi/ml, 10 µM) supplemented with D1i (1 µM) or D2i (10 µM), in presence or absence of 100 µM oleic acid for 4 h (A,D) or with 100 µM [¹⁴C]acetate (2 µCi/ml) in presence or absence DGAT inhibitors for 4 h (B,E). C,F) Cells were incubated with the liver X receptor agonist T0901317 (1 µM) for 96 h. Thereafter, myotubes were incubated for 24 h with D-[¹⁴C(U)]glucose (2 µCi/ml, 5.5 mM) in presence or absence of DGAT inhibitors. Lipids were separated by thin layer chromatography and measured using liquid scintillation. The sum of all radioactivity on the TLC plate is presented as total lipids. Results represent mean ± SEM from n = 4 (A,B), n = 5 (D,E) or n = 3 (C,F) individual experiments with 3 separate culture wells for each condition presented at absolute values (A, B, C) or normalized to control (D–F). *p < 0.05 vs. control, paired t-test. D1i, DGAT1 inhibitor; D2i, DGAT2 inhibitor; TAG, triacylglycerol.