Figure 2

Effect of DGAT inhibition on lipid droplets and total cellular triacylglycerol content. Human myotubes were grown and differentiated on glass bottom microwell dishes. At day 7 of differentiation, myotubes were incubated with 100 µM OA for 4 (A–C) and 24 h (D) in presence or absence of DGAT inhibitors; D1i (1 µM) and D2i (10 µM). The cells were stained for lipid droplets (green) and nuclei (blue) using Bodipy 493/503 and Hoechst 33528, respectively, and images taken with a 60x objective on a confocal microscope. Scale bar 25 µm. (A–C) Representative images are presented for control (A), D1i (B) and D2i (C). (D) LDs were quantified (ImageJ) by relating number of LDs to number of nuclei. Results represent mean ± SEM from one experiment at 4 h and one experiment at 24 h where calculations are based on 6–8 different images for each condition, unpaired t-test, *p < 0.05 vs control at the same time-point. (E) Human myotubes were incubated with 100 µM OA in presence or absence of D1i (1 µM) and D2i (10 µM) for 24 h and total TAG content was measured. The results are presented as mean ± SEM as % of control from n = 3 individual experiments with one 75 cm² cell culture flask for each condition. *p < 0.05 vs. control, paired t-test. D1i, DGAT1 inhibitor; D2i, DGAT2 inhibitor; LDs, lipid droplets; OA, oleic acid; TAG, triacylglycerol.