FIGURE 4:

Overexpression of ATP9A affects Tfn trafficking. HeLa cells were transiently transfected with expression vector encoding HA-tagged ATP9A (A) or ATP9B (B), serum starved for 3 h, and then incubated at 4°C for 50 min with Alexa Fluor 488–conjugated Tfn. Cells were washed and chased at 37°C for 5 min and 20 min in the absence of the labeled Tfn. Cells were fixed and incubated with anti-HA antibody followed by Cy3-conjugated anti-rat secondary antibody (asterisks: overexpressing cells). Scale bars: 20 μm.