FIG. 3.

Infected cells treated with geldanamycin show reduced viral DNA production and virus yield. (A) Viral DNA synthesis is impaired in cells treated with Hsp90 inhibitors. Total viral DNA isolated from infected cells at 6 and 18 h postinfection was subjected to PFGE and Southern blotting. Samples include uninfected cells (lanes 1 and 2), HSV-1-infected cells (lanes 3 and 4), HSV-1-infected cells treated with GM (lanes 5 and 6), and HSV-1-infected cells treated with a derivative of GM, 17AAG (lanes 7 and 8). Newly replicated viral DNA molecules (“Well”) are large, probably branched molecules that fail to enter the pulsed-field gel. Unit-length molecules (152 kb) are readily observed at 18 h postinfection in wild-type HSV-1 infection (lane 4). DNA synthesis is severely impaired in cells treated with either GM or 17AAG (lanes 5 to 8). No signal is observed in the mock-infected samples. (B) Virus yield is reduced in cells treated with geldanamycin. HSV-1-infected Vero cells were incubated in the absence (black bar) or presence (dark gray bar) of 1.0 μM geldanamycin for 6 or 24 h. Infections were performed in triplicate. Virus titers were determined by plaque assay as described in Materials and Methods.