Figure 5.
Reovirus Replication Is Enhanced by KU60019 Treatment
(A) Supernatants of reovirus (MOI 100 for CMeC1, KMeC, and LMeC, or MOI 10 for CMGD2)-infected cell lines treated with KU60019 (2.5 μM) were harvested after a 48-hr incubation, and virus titers were determined by the TCID50 assay. The fold increase of reovirus titer represents values calculated from the titer of progeny virus divided by the titer of input virus. The mean + SD was calculated from three independent experiments. p values were calculated by Tukey-Kramer test, *p < 0.05, **p < 0.01; ns, not significant. (B) Effects of ribavirin (200 μM) in CMeC1 treated with combination of reovirus (MOI 100) and KU60019 (2.5 μM) for 24 hr were assessed by western blotting using anti-reovirus antibody and anti-cleaved caspase-3 antibody. Actin was used as a protein loading control. (C and D) CMeC1 cells were pre-treated with reovirus virions (C, MOI 100) or ISVPs (D, MOI 10) and washed-out unattached virus followed by treatment with KU60019 (2.5 μM) for the indicated time points. Reovirus structural proteins were detected by western blotting using anti-reovirus polyclonal antibody. Actin was used as a protein loading control. The relative level of reovirus protein normalized to actin was quantified from at least three independent experiments. Mean + SD are shown. Tukey-Kramer test, *p < 0.05, **p < 0.01.