Figure 6.

Knockdown of the endogenous ATF6 gene inhibits aMPV/C-induced autophagy and virus production. (A) Vero cells were mock-transfected (mock) or transfected with ATF6-siRNA at different concentrations (10, 20, 30 or 40 pmol), or 40 pmol scrambled-siRNA (Scra). ATF6 protein was detected by western blotting at 48 h. (B) Vero cells were transfected with ATF6-siRNA, Scra and Con (siRNA-untransfected) and then infected with aMPV/C for 72 h. The cells were harvested and analyzed by western blotting with anti-ATF6, anti-SQSTM1, anti-LC3, anti-viral F, and anti-ACTB antibodies. Representative results are displayed with graphs corresponding to the ratios of ATF6:ACTB or viral F:ACTB normalized to the control conditions. (C) Vero cells were transfected and infected as described in B. Progeny virus yields were examined by TCID₅₀ at 72 hpi in Vero cells. (D) Vero cells were transfected with pCMV-HA (HA) or pCMV-HA-ATF6 (HA-ATF6) plasmid and then infected with aMPV/C for 72 h. The cells were harvested and analyzed by western blotting with anti-ATF6, anti-SQSTM1, anti-LC3, anti-viral F, and anti-ACTB antibodies. Representative results are displayed with graphs corresponding to the ratios of ATF6:ACTB or viral F:ACTB normalized to the control conditions. (E) Vero cells were transfected and infected as described in D. Progeny virus yields were examined by TCID₅₀ at 72 hpi in Vero cells. Error bars, mean ± SD of 3 independent experiments. Two-way ANOVA test, ^#P > 0.05; *P < 0.05; ***P < 0.001, compared with the control group.