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Prohibitin plays a critical role in Enterovirus 71 neuropathogenesis

Fig 3

siRNA-mediated gene silencing in NSC-34 cells.

NSC-34 cells were transfected with various concentrations of ON-TARGET PLUS siRNA SMARTpools. (a) At 48 h.p.t., the cell viability of transfected cells was assessed using alamarBlue assay. (b) Knocked-down NSC-34 cells were infected with EV71 at M.O.I. 10. The viral titers in the culture supernatant were determined by plaque assay at 48 h.p.i.. Statistical analysis was performed using one-way ANOVA test with Dunnett’s posttest (* p<0.1, ** p<0.01, *** p<0.001, **** p<0.0001). (c) NSC-34 cells were transfected with PHB siRNA pool or with non-targeting siRNA (NTC) control at various concentrations. The efficiency of siRNA knockdown was verified at 48 h.p.t. by Western blot. (d) PHB siRNA- or siNTC-transfected cells were infected with EV71 at M.O.I. 10. The viral titers in the culture supernatants were determined by plaque assay at 48 h.p.i.. Cell viability of the transfected cells was assessed using alamarBlue assay. Statistical analysis was performed using two-tailed student’s t-test (** p<0.005, *** p<0.005). Relative band quantification (below Western blot) was determined by ImageJ, by normalizing to loading control, β-actin. Error bars represent mean ± standard deviation. One representative of two biological repeats is shown.

Fig 3

doi: https://doi.org/10.1371/journal.ppat.1006778.g003