Fig. 7. Coexpression of HA-GLUT4s and dominant negative dynamin. Isolated cells were cotransfected with 0 (–) or 12.5 (+) µg/ml dynamin-K44A plasmid/cuvette and 2.5 µg/ml of expression plasmid for HA-GLUT4s and cultured for 24 hours. After harvesting, the cells were incubated without (basal, solid bars) or with (open bars) 67 nM insulin for 30 minutes, and the cell-surface levels of HA-GLUT4s were determined using an antibody binding assay as described in Materials and Methods. The cell surface-associated radioactivity was normalized to the relative protein expression level of each respective mutant. Results are the means of the mean values (
,
) obtained from at least duplicate determinations in two independent experiments.
