FIG 6.

Reovirus transcripts are present in viral factories only when σNS is capable of binding RNA. Cells that constitutively express siRNAs directed against σNS were transfected with expression plasmids encoding the σNS constructs shown and incubated for 24 h. Cells were adsorbed with reovirus strain T3D at an MOI of 5 PFU/cell, incubated for 48 h, fixed, stained using RNA FISH probes specific for σNS mRNA (white) or σ3 mRNA (yellow), μNS-specific antiserum (magenta), and DAPI (blue), and imaged using confocal microscopy. Bar, 4.8 μm. The percentage of cytoplasmic σNS-mRNA (B, white bars) and σ3-mRNA (B, yellow bars) FISH signals in reovirus factories was quantified by dividing the sum of the FISH signal in reovirus factories by the sum of the cytoplasmic FISH signal. Individual data points represent single cells. Percentage values that differ significantly from those obtained from WT σNS-transfected reovirus-infected cells by one-way analysis of variance (ANOVA) with Dunnett’s multiple-comparison test are shown. *, P < 0.0332; ****, P < 0.0001.