Figure 6

The effect of NS1 NES mutations on recombinant virus-induced interferon production and IRF3 and Akt phosphorylation. A549 cells on 6-well plates were infected at three different MOI values for 20 h after which the supernatants and cell lysates were collected. A and B, quantitation of IFN-β and IFN-λ1 from the supernatants by ELISA. All samples were analyzed in duplicates and the data shown is representative of three independent experiments. Statistical significance is calculated against the wt samples (*p < 0.05, **p < 0.01). C, Virus-infected cells (MOI 1) were collected at 20 h postinfection, lysed and processed for SDS-PAGE and Western blotting in three different dilutions (10, 5 or 1 μg of total protein/well). Filters were stained with antibodies against p-IRF3, p-Akt, MxA, actin and NS1 proteins and relative protein expression is shown in the figure. Data shown is representative of three independent experiments.