FIG. 4.

AAA-PKB inhibits insulin-induced translocation of GLUT4myc in muscle cells. L6-GLUT4myc myoblasts were cotransfected with GFP and HA-AAA-PKB (0.4 μg each) and incubated for 48 h in culture. Cells were serum deprived for 5 h, left untreated or treated with 100 nM insulin for 20 min (insulin), and then processed for cell surface GLUT4myc detection with anti-myc (1:100) antibody followed by Cy3-conjugated goat anti-mouse antibody as indicated in Materials and Methods. Each pair of panels (upper and lower) shows the same field of cells. In the lower panels, GFP fluorescence in transfected cells is shown (arrowheads). The upper panels show the cell surface GLUT4myc density under basal conditions (a) or with insulin treatment (b) for three separate experiments. Arrowheads indicate the positions of cells transfected with HA-AAA-PKB in the upper panels. Results shown are representative of at least five experiments.