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. 2004 Dec;78(23):12747–12761. doi: 10.1128/JVI.78.23.12747-12761.2004

FIG. 2.

FIG. 2.

FIG. 2.

FIG. 2.

Higher induction of VSV-mediated apoptosis in PERK−/− MEFs. PERK+/+ (A) and PERK−/− (B) MEFs were left untreated or treated with either mouse IFN-α/β (1,000 IU/ml) or IFN-γ (100 IU/ml) for 20 h, followed by infection with VSV at an MOI of 1. Cells were harvested at 24 or 36 hpi and subjected to annexin V-PI staining (BioSource) according to the manufacturer's specifications. Cells were then subjected to flow cytometry analysis by using FACScan (Becton Dickinson), and data were analyzed by using WinMDI version 2.8 software (The Scripps Institute). Cells were gated on a dot plot showing forward and side scatter in order to exclude debris not within the normal size. Gated cells were plotted on a dot plot showing annexin V staining (FL1-H) and PI straining (FL2-H). The numbers represent the percentage of gated cells counted for their corresponding quadrant. These are data of one out of three reproducible experiments. (C and D) PERK+/+ and PERK−/− MEFs were left uninfected or were infected with VSV at an MOI of 1 (C) or an MOI of 100 (D); protein extracts (30 μg) were subjected to immunoblot analysis with an anti-VSV antibody (right panels), or virus titers were measured by harvesting medium at the indicated times postinfection, followed by plaque assay analysis (left panels). Symbols: ▴, virus titers from PERK+/+ MEFs; ▪, virus titers from PERK−/− MEFs.