Fig. 2. Cell surface expression of HA-GLUT8 in HeLa cells. Cells were grown in six-well plates and transfected with HA-GLUT8 without or with plasmid for dynamin K44A, and with mutant HA-GLUT8-LL/AA. After 48 hours, cells were harvested and the expression of the HA-GLUTs was determined by western blotting using an anti-HA antibody. In parallel, cell-surface levels of the HA-GLUTs were determined using an antibody binding assay as described in the Materials and Methods. The cell surface-associated radioactivity was normalized to the relative protein expression level of each respective mutant. Results are the means ± s.d. of a representative experiment performed in duplicate.