Prohibitin plays a critical role in Enterovirus 71 neuropathogenesis
NSC-34 cells were first infected with EV71 at M.O.I. 10, followed by Roc-A treatment for 48 hours. At 48 h.p.i. the culture supernatant was collected for viral titer determination by plaque assay (a). Furthermore, the cell lysate was prepared for Western blot analysis (b). Relative band quantification (below Western blot) was determined by ImageJ, by normalizing to loading control, β-actin. Cell viability was assessed using alamarBlue assay. (c) Immunostaining of Roc-A treated NSC-34 cells. At 48 hours incubation the cells were fixed with ice cold methanol and probed with specific antibodies. Scale bar denotes 20 μm. (d) Mitotoxicity of Roc-A in NSC-34 cells. Cells were incubated with various concentrations of Roc-A for 48 hours before assessment of cytotoxicity (fluorescence) and mitotoxicity (luminescence) using Mitochondrial ToxGlo Assay. Statistical analysis was performed using one-way ANOVA with Dunnett’s post-test (*, p<0.05; **, p<0.005; ***, p<0.001; ****, p<0.0001). One representative from two independent experiments is shown.