FIG. 6.

siRNA directed against eIF4E inhibits VSV replication in primary activated CD4⁺ T lymphocytes. (A) Primary CD4⁺ T lymphocytes were stimulated with anti-CD3/CD28 for 48 h before electroporation with eIF4E-specific siRNA or scrambled siRNA. After an additional 48-h incubation period, cells were lysed and analyzed by immunoblotting with an antibody against total eIF4E. “−” denotes cells electroporated without siRNA. (B) Cell cycle analysis was performed on cells described in legend to panel A by PI-labeling and flow cytometry analysis. In panels B and C, the percentages of cells in various phases of the cell cycle are indicated. The data shown represent the means ± standard deviations (n = 3). Statistically significant differences are indicated by asterisks (P < 0.005). (D) eIF4E knockdown and control cells were infected with VSV (1 MOI) for 12 h. Cells were lysed and subjected to immunoblotting with anti-VSV antisera. The eIF4E and VSV N/P protein expression levels in panels A and B were quantified and normalized to β-actin levels using the Scion Image 4.0 software program. Levels of eIF4E were expressed as percentages of levels for mock-infected cells.