Fig. 5. Effects of HA-GLUT4-Akt2 fusion proteins on cell-surface expression of myc-GLUT4 in rat adipose cells. Isolated cells were co-transfected with myc-GLUT4 and either nothing, HA-GLUT4-Akt2-wt, HA-GLUT4-Akt2-S474A, or HA-GLUT4-Akt2-KD, and cultured for 20 h. After harvesting the cells, cell-surface levels of myc-GLUT4 were determined in the transfected cells in the basal (white) and insulin-stimulated states (67 nM, black) using a myc antibody binding assay, and all of the values were expressed relative to the mean basal myc-GLUT4 value in each experiment as described in Materials and Methods. Results are the means ± s.e.m. of the means obtained from at least duplicate determinations in each of 3-5 independent experiments.