FIG. 8.

Recycling of internalized GFP-gB and LL871AA to the plasma membrane. After transfection, GFP-gB- or LL871AA-expressing cells were stained with an anti-GFP antibody at 4°C and then either immediately fixed (A and D) or incubated for 20 min at 37°C to allow internalization (B, C, E, and F). After incubation, cells were subjected to an acid-glycine treatment and then immediately fixed under nonpermeabilizing conditions (B and E) or placed again at 37°C for 30 min before fixation (C and F). Fixed cells were labeled with a Cy3-coupled secondary antibody and examined by confocal microscopy. Insets show the spontaneous GFP fluorescence emitted in transfected cells.