FIG 7.

vhs-GFP expressed in virus infection is deficient in endoribonuclease activity. (A) HFFF cells were infected with WT or HSV1 vhs-GFP viruses at an MOI of 5. Total RNA was purified at 16 hpi and subjected to RT-qPCR for cellular transcripts previously identified as being susceptible to vhs activity. The mean and ± standard error for n = 3 is shown. Statistical analysis was carried out using an unpaired t test. ***, P < 0.001; ****, P < 0.0001. (B) RNA samples from (A) were subjected to RT-qPCR of viral transcripts ICP22 (IE), TK (E), and gC (L). The mean and ± standard error for n = 3 is shown. Statistical analysis was carried out using an unpaired t test. ***, P < 0.001; ****, P < 0.0001; ns, not significant. (C) HFFF cells infected with WT or HSV1 vhs-GFP viruses at MOI of 2 were harvested at 16 h and analyzed by SDS-PAGE and Western blotting with antibodies as indicated. Western blotting for eIF2α was carried out using 10 μm Phos-tag PAGE. The relative level of phospho-PKR and phospho-eIF2α was quantitated using LI-COR Image Studio and is represented relative to the level in uninfected cells.