Figure 5.

rabip4′ FYVE domain does not affect endosome morphology and Tf recycling. Pulse-chase experiments in control HeLa cells (A and B) or HeLa cells transfected with HA-rabip4′-FYVE (C-D′) or GFP-FENS1-FYVE (E-F′). Cells expressing HA-rabip4′-FYVE were labeled with mouse anti-HA tag followed by Alexa488-goat anti-mouse IgG (C and D), whereas expression of GFP-FENS1-FYVE was visualized by the GFP signal (E and F). Alexa594-Tf was internalized for 30 min at 16°C, washed, and chased at 37°C for 0 min (A, C′, and E′), and 20 min (B, D′, and F′) in the presence of 50 mM Desferal. Arrows denote colocalization of Alexa594-Tf and GFP-FENS1-FYVE. Bar,10 μm.