FIG 15.

EBOV vncRNAs are not associated with the RISC and do not positively or negatively affect virus replication. (A) qRT-PCR analysis of EBOV vncRNAs following RNA-binding protein immunoprecipitation of AGO1 to -4 with individual AGO antibodies and a pan-AGO antibody. Combined data from two independent experiments are shown; the data for pan-AGO RIP are representative of five separate experiments. SAM-68 was used as an irrelevant RNA-binding protein control. Of the four EBOV vncRNAs profiled, only data for the EBOV GP vncRNA are shown. (B) EBOV titers in supernatants of 293T-P cells at 48 hpi. Prior to infection, cells were transfected with the indicated total concentrations of siRNAs homologous to the VP40 and L vncRNAs. siRNAs were transfected individually or combined (combined concentrations, 12.5 nM each or 25 nM each). Scrambled control siRNA-transfected or mock-transfected cells were used as controls for comparison. Significance was tested by two-way analysis of variance (α = 0.05) followed by Dunnett’s multiple-comparison posttest, and separate independent comparisons were made to the mock-transfected and scrambled siRNA-transfected controls. The means for three biological replicates ± standard deviations are plotted.