Fig. 1.
Myc-adipose triglyceride lipase (ATGL) has intact triacylglycerol (TG) hydrolase activity. A: schematic structure of Myc-tagged mouse ATGL with functional domains. B: extracts of HeLa cells transfected with control (Ctrl) vector, ATGL, Myc-ATGL, or comparative gene identification-58 (CGI-58) were mixed in various combinations. TG hydrolase activity was measured using 3H-labeled triolein as substrate and normalized with the total protein levels of the cell extracts. Data are shown as means ± SD and represent 3 independent experiments.*P < 0.05; **P < 0.01; ***P < 0.001. Expression of ATGL, Myc-ATGL, and CGI-58 were analyzed by anti-ATGL/CGI-58 immunoblotting, using β-actin as a loading control. C: HeLa cells transfected with Myc-ATGL were incubated under growth conditions with 400 μM oleic acid complexed to albumin for 16 h. Immunofluorescence staining with anti-Myc antibody (red) was performed to reveal Myc-ATGL. Lipid droplets (LDs) were costained with Bodipy 493/503 fluorescence dye (green).