FIG. 7.

Effect of Sam B on HSV-1 DNA synthesis in Vero cells detected by PCR. Cells (5 × 10⁶) were infected with HSV-1 (MOI, 3) or were not infected in the presence or absence of Sam B (25 μM). The cells were harvested at 2, 4, 8, and 16 h p.i., and total DNA was extracted with phenol-chloroform. The PCR was done as described in Materials and Methods. Following the reaction, the amplified product was run on a 2% agarose gel. (A) DNA was extracted from uninfected Vero cells at 16 h p.i. (lane 1) or from HSV-1-infected cells at 2, 4, 8, and 16 h p.i. (lanes 2 to 5). (B) Sam B and HSV-1 were added to the cells at the same time, and the total cellular DNA was extracted at 2 (lane1), 4 (lane 2), 8 (lane 3), and 16 h (lane 4) p.i. Lane 5, total cellular DNA extracted from infected cells treated with Sam B at 12 to 16 h p.i. Each band was quantitated by densitometer, and the ratio of HSV-1 DNA to β-actin DNA was calculated. Each bar represents the mean of three independent experiments.