Fig. 3

ETC inhibition stongly suppresses viral replication. Cells were infected by MHV (MOI 0.05) for 24 h. Chemicals were treated after serum free medium is changed to FBS containing medium. A, D, G RNA expression of MHV in cells treated with ETC inhibitors (Oligomycin, Omy; Rotenone, Rot; FCCP) in replete medium (glucose 450 mg/dl, pyruvate 0.5 mM). Rplp0 was used for an internal control. B, E, H Western blot analysis and quantification of SPIKE in cells treated with ETC inhibitors (Omy, Rot, FCCP) in replete medium (glucose 450 mg/dl, pyruvate 0.5 mM). β-actin was used for an internal control. C, F, I Viability of cells incubated for 24 h with medium collected from MHV-infected cells which were treated as indicated. J RNA expression of MHV in cells pre-treated with ETC inhibitors (indicated doses) 1 h before MHV infection. Cells were washed and harvested in 0.5 hpi. Rplp0 was used for an internal control. K Illustration of metabolic intervention induced by ETC inhibition. Values represent means ± SD. *p < 0.05. One-way ANOVA followed by a tukey’s multiple comparison test was performed. All experiments were repeated at least 3 times