FIG. 5.

Inhibition of viral DNA synthesis reduces the late expression of IE genes in wild-type and UL21a deletion virus-infected cells to equivalent levels. (A) Analysis of viral protein accumulation in the presence of PAA. MRC-5 cells were infected with ADwt or ADsubUL21a at an input genome number equivalent to 3 TCID₅₀ units of wild-type virus/cell in the presence or absence of PAA (100 μg/ml). Cells were collected at the indicated times, and total cell lysate was analyzed by immunoblotting with the indicated antibodies. (B) Analysis of viral transcript accumulation. MRC-5 cells were infected in the presence (+) or absence (−) of PAA as for panel A, total RNA was collected, and selected IE transcripts were analyzed by RT-qPCR and normalized to GAPDH. The normalized amount of viral transcript at 24 hpi during wild-type virus infection was set to 1. n.s., P > 0.05; *, P < 0.05 (by Student's t test). Error bars indicate standard deviations. (C) Viral protein accumulation in the presence of PAA or ganciclovir (GCV) during infection of HCMV strains AD169 and TR. MRC-5 cells were infected with ADwt (left panel) or TR (right panel) at an MOI of 2 with addition of no drug, PAA (100 μg/ml), or GCV (30 μg/ml). Cells were harvested at the indicated times and analyzed by immunoblotting with the indicated antibodies. Shown are representative results from at least two independent experiments.