Fig 2.

Colocalization of wild-type or mutant NS1 proteins with overexpressed RAP55 and suppression of RAP55-associated stress granule formation by different versions of NS1. HEK 293T cells were cotransfected with V5-tagged RAP55 and wild-type and mutated Flag-tagged NS1 expression vectors (NS1 mutants are shown in Fig. 1B). Signals were examined by double immunostaining using anti-Flag and anti-V5 antibodies at 16 to 20 h posttransfection. Formation of stress granules was detected using an anti-mouse monoclonal antibody specific for endogenous G3BP1. (B) Growth kinetics of WSN wild-type and mutant (R35A and I123/A/M124A/K125A/M127A) viruses. A549 cells were infected with WSN WT, R35A mutant, or I123/A/M124A/K125A/M127A mutant virus at an MOI of 0.05. Viral titers were determined by plaque assay at the indicated time points. Values represent the means for three independent experiments, and error bars show standard deviations. *, P < 0.05; **, P < 0.005.