FIG. 3.

Colocalization between gB and α-adaptin in gB-transfected cells. PK15 cells were transiently transfected with either Be gB (pGVM3), Y864A gB (pGVM16), Y902A gB (pGVM17), or gB truncated (pGVM50). At 24 h posttransfection, the cells were incubated with FITC-labeled monoclonal anti-gB antibodies at 4°C for 1 h. Subsequently, the cells were shifted for 60 min to 37°C to allow internalization of gB (right column). Thereafter, the cells were fixed, permeabilized, and costained for α-adaptin (middle column) by the use of monoclonal anti-α-adaptin IgG1 antibodies, biotinylated IgG1-specific antibodies, and streptavidin-Texas Red. Merged images (right column), obtained by superposition of the green (gB) and red (α-adaptin) stainings, indicate areas of colocalization (yellow) between gB and α-adaptin. The arrows indicate colocalization between wild-type or Y864A gB and α-adaptin in cytoplasmic internalization vesicles. Bar, 10 μm.