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. 2011 Feb 2;6(2):e16711. doi: 10.1371/journal.pone.0016711

Figure 4. Anti-PKR mechanisms can be exchanged between viruses.

Figure 4

A) Previous infection with some viruses prevented eIF2α phosphorylation after SINV virus superinfection. The protocol followed for mixed infections is outlined. Wild type MEFs were infected with the indicated viruses at a moi of 2 pfu/cell. Three hours later, cells were superinfected with SINV at moi of 25 pfu/cell and 5 h later lysed in sample buffer for immunoblot analysis against SINV capsid (SINV C, upper panel) and anti-phosphoeIF2α (bottom panel). B) Expression of VV E3L gene rescued translation of ΔDLP SINV mutant. MEF-E3L cells were induced for the expression of VV E3L by tetracycline withdrawal and infected with SINV-WT or SINV-ΔDLP mutant. Five hours later, cells were labeled with [35S]-Met+Cys for 30 min and analyzed by SDS-PAGE followed of autoradiography and by immunoblot against anti-E3, anti-PKR and anti-phospho eIF2α. Parallel cultures were infected at moi of 5 pfu/cell and viral yields were determined by plaque assay 2 days later. Data are the mean ±SD from three and two independent experiments in PKR+/+ and PKRo/o cells, respectively.