Figure 4.

Effect on DGAT inhibition on turnover of accumulated lipids. Human myotubes were grown and differentiated on 96-well tissue culture plates, SPA plates were used for time-course (B,C). At day 6 of differentiation myotubes were incubated with [¹⁴C]oleic acid (0.5 µCi/ml, 100 µM) for 24 h. After 24 h pre-treatment with [¹⁴C]oleic acid myotubes were washed and re-incubated with D1i (1 µM) or D2i (10 µM). (A) Cell-associated radioactivity from [¹⁴C]oleic acid was measured after 4 h treatment with DGAT inhibitors. (B,C) Decline in cell-associated radioactivity was measured over 6 h in presence of an inhibitor of carnitine palmitoyltransferase 1 (etomoxir, 10 µM) and DGAT inhibitors; D1i (1 µM) and D2i (10 µM). Results represent mean ± SEM as nmol/mg protein (A,B) and as all-over effects normalized to control (C) from n = 4 individual experiments with 8 separate culture wells for each condition. *p < 0.05 vs control, paired t-test (A), LMM statistical test (C). D1i, DGAT1 inhibitor; D2i, DGAT2 inhibitor; SPA; scintillation proximity assay; LMM, Linear mixed model.