Figure 1
Effect of siRNA treatment targeting the 5’-UTR on PV replication.
(a) Relative expression of PV in Vero cells transfected with 13 different siRNAs was detected using real-time PCR. All data are shown as the mean ± standard deviation based on three independent experiments and displayed as the fold change over the siRNA-NC controls. (b) Confocal microscopy analysis of PV in PV-infected cells. Vero cells were transfected with siRNA-NC or siRNA-100 before PV infection, as described in Experimental Procedures. Immunostaining for PV (red) was performed 24 h later using mouse anti-PV at 4 °C overnight and goat anti-mouse secondary antibody conjugated to rhodamine (TRITC). The cells were counterstained with Hoechst 33258 (nuclear, blue) and photographed using a Nikon confocal microscopy system. PV was expressed at high levels in virally infected cells transfected with siRNA-NC but at low levels in siRNA-100-treated cells. Typical nuclear CPE was observed in siRNA-NC-treated cells but not in siRNA-100-treated cells. Bars represent 20 μm. Vero cells (c) and A549 cells (d) with or without siRNA transfection were infected with PV at a TCID50 of 10−6. The cells and supernatants were harvested at different times post-transfection and the total virus titers were determined by a TCID50 assay.
