FIG. 4.

(A) Viral response to IFN. Monolayers of Vero cells were either untreated or pretreated with IFN-α (200 U/ml; Sigma) for 20 h. Cells were then infected with indicated viruses at 0.05 PFU per cell and incubated at 37°C. At 24 h postinfection, cells were harvested and freeze-thawed three times, and virus yields were determined by titration. (B) Synthesis of viral proteins and eIF2α dephosphorylation. Monolayers of HEL fibroblasts were either mock infected or infected with viruses as indicated (5 PFU/cell). At 18 h after infection, lysates of cells were subjected to electrophoresis and reacted with antibodies against mixed HSV-1 antigens (Dako Coporation), eIF2α, and phosphorylated eIF2α (Cell signaling Tech). Size markers are listed on the left. (C) Viral induction of antiviral genes. HEL fibroblast cells were mock infected or infected with HSV-1(F), R3616, or H9813 (5 PFU/cell) as indicated. At 6 h postinfection, total RNA was extracted from cells and subjected to reverse transcription-PCR amplification and electrophoresis for ISG56, ISG54, and 18s rRNA.