FIG 5.

Mutations in σNS that compromise RNA binding disrupt incorporation of the protein into factory-like structures. (A) Cells were transfected with expression plasmids encoding the σNS constructs shown along with μNS and incubated for 24 h. Cells were fixed, stained using σNS-specific monoclonal antibody 3E10 (cyan), μNS-specific antiserum (magenta), and DAPI (blue), and imaged using confocal microscopy. Regions selected for magnification are indicated by dotted white boxes. Bar, 5 μm. (B) The percentage of σNS immunofluorescence signal in reovirus factory-like structures was quantified by dividing the sum of σNS signal in reovirus factory-like structures by the sum of cytoplasmic σNS signal. Individual data points represent single cells. Percentage values that differ significantly from those obtained from WT σNS and μNS cotransfected cells by one-way analysis of variance (ANOVA) with Dunnett’s multiple-comparison test are shown. ****, P < 0.0001.