Fig. 9. Co-staining of HA-GLUT8, HA-GLUT8/YTRF and transferrin in HeLa cells. Cells were grown on coverslips and either transfected with HA-GLUT8 or HA-GLUT8/YTRF. After 3 days in culture, serum-starved cells were incubated with transferrin (Tf) conjugated to Alexa Fluor 633 for 30 minutes at 37°C, thoroughly washed with PBS and fixed in 4% paraformaldehyde. Then the cells were stained for the HA-epitope tag and analyzed by confocal laser scanning microscopy as described in the Materials and Methods Squares indicate magnified regions (mag.). Bars, 20 µm.