FIG. 2.

Confocal microscopy analysis of gB internalization in KOS-infected cells. 143B cells were infected with wild-type KOS virus at an MOI of 0.1. At 7 h postinfection, the cells were incubated with an anti-gB antibody at 4°C for 45 min and then placed at 37°C for 0 min (A, E, I, M, and Q), 15 min (B, F, J, N, and R), 30 min (C, G, K, O, and S), or 60 min (D, H, L, P, and T). Cells were fixed in methanol and labeled with anti-TGN 46 and anti-VP5 antibodies. Panels A to D correspond to phase-contrast images of infected cells, whereas panels E to P show indirect immunofluorescence from anti-VP5 and Cy2-labeled secondary antibodies (green) (E to H), from anti-TGN 46 and Cy5-labeled secondary antibodies (pseudo-colored in blue) (I to L), or from anti-gB and Cy3-labeled secondary antibodies (red) (M to P). (Q to T) Merged images. Fluorescence was visualized with a Leica TCS SP2 AOBS confocal microscope.