FIG 3.

Mutants of σNS incapable of RNA binding fail to complement reovirus replication in cells expressing σNS-specific siRNAs. (A) Cells that constitutively express siRNAs directed against GFP or σNS were transfected with expression plasmids encoding GFP or the σNS constructs shown and incubated for 24 h. Cells were adsorbed with reovirus strain T3D at an MOI of 5 PFU/cell and incubated for 48 h. (B) Cell culture supernatants were collected for infectious virus quantification by plaque assay. Titer values that differ significantly from those obtained from cells expressing siRNAs against GFP complemented with GFP by one-way ANOVA with Dunnett’s multiple-comparison test are shown. **, P < 0.0021; ***, P < 0.0002; ****, P < 0.0001. (C) Cells that constitutively express siRNAs against GFP or σNS were transfected with expression plasmids encoding the σNS constructs shown and incubated for 24 h. Cell lysates were resolved by SDS-PAGE and immunoblotted using antisera directed against σNS and monoclonal antibodies specific for alpha-tubulin (α-Tub). Panel A was prepared using BioRender.