Figure 6.

Confocal microscopy of VV-infected cells. (A–B) Microtubules are needed for IEV transport to the cell periphery. RK₁₃ cells were infected with VV at 1 PFU/cell for 11.5 h. The medium was replaced by either fresh DME (A) or DME containing 33 μm nocodazole (B). At 13 hpi, the cells were fixed, permeabilized, and stained with Mab 19C2 (anti-B5R). Arrows indicate clusters of virus particles near the cell surface. (C–F) PP1 blocks the formation of actin tails but not CEV particles. RK₁₃ cells were infected with VV at 1 PFU/cell, and at 5 hpi PP1 was added to a final concentration of 20 μM. (C and D) At 11 hpi, the medium was replaced with DME containing mAb 19C2 (anti-B5R) (live cells), and the cells were incubated for 1 h before fixation and processing for fluorescent microscopy. (E and F) Cells were fixed at 12 hpi and stained with and rhodamine-conjugated phalloidin. Bars, 10 μm.