Figure 6.

Reduced transport of capsids to the nucleus in the absence of MTs. Immunofluorescence microscopy of Vero cells infected with HSV-1 for 2 h at an MOI of 50 in the presence of CH and drugs affecting the cytoskeleton. (a) Cytoskeletal drugs. Under control conditions, almost all capsids accumulate around the nucleus. If the cells were infected in the presence of nocodazole, colchicine, or vinblastine, respectively (lower panel), there is no accumulation of viral capsids at the nucleus; instead, the capsids are scattered throughout the entire cytoplasm. In cells infected in the presence of taxol or cytochalasin D, the capsids accumulate at the nucleus (upper panel). The cells were fixed in 3% PFA, permeabilized with 0.1% TX-100, and labeled with an antibody directed against purified capsids (HC). (b) Reversibility. In the presence of nocodazole, the amount of viral capsids localized to the nucleus is drastically reduced, whereas, after a 2-h chase in nocodazole-free medium, the capsids accumulate around the cell nuclei. Cells were infected in the presence of nocodazole for 2 h, fixed immediately (left panels), or washed and further incubated in normal medium for an additional 2 h before fixation (right panels). After extraction with 0.5% TX-100 in MT buffer and fixation in methanol, the cells were double labeled with anti-tubulin (left panels) and anti-capsid (HC, right panels).