Figure 5.

Rapid recycling from early endosomes is impaired by CBD2 overexpression. tTA HeLa cells were infected with control or AAK1L CBD2 encoded adenovirus for 24 h. Infected cells were incubated for 1 h at 16°C with 5 μg/ml biotinylated transferrin (BXX-Tfn) and acid washed to remove uninternalized ligand. Early endosome recycling was then scored at the indicated time points after transfer to 37°C (A). Alternatively, cells were incubated at 37°C for 10 min to allow Tfn transport to the endosome recycling compartment, acid washed again, and then scored for recycling (B). Error bars, ±SD of 3–4 independent experiments.