Fig 3.

dsRNA-triggered eIF2α phosphorylation and cell death were alleviated by JEV NS2A. (A) Stable A549 cells transduced with lentiviral vectors expressing HA-tagged GFP, JNS2A (J2A), and PKRΔ6 were stimulated with poly(I·C) (0, 0.4, 1, and 2 μg/ml) for 1 h. The cell lysates were then harvested for Western blot analysis with the indicated antibodies. The bands intensities were quantified by MetaMorph (Universal Imaging Corp.), and the relative levels for the indicated proteins are shown. (B) The parental, stable GFP-expressing, and J2A-expressing A549 cells described in panel A were stimulated with poly(I·C) (0, 5, and 10 μg/ml) for 8 h. Viable cell numbers were determined by the trypan blue exclusion method. The relative cell survival of poly(I·C)-treated cells was compared to that without poly(I·C) treatment for each type of cell. The experiment was performed three times with similar results, and representative data from one experiment performed in triplicate are shown with means and the standard deviations (n = 3). **, P < 0.005; ***, P < 0.001 (two-tailed Student t test).