Fig. 5

Inhibition of autophagy with specific siRNA targeting ATG5 reduces CDV replication. A and C Vero cells were transfected with siRNA-ATG5 or scrambled siRNA for 48 h; then, the cells were infected with CDV (MOI = 1). The cell samples were then analyzed by western blot with anti-ATG5, anti-CDV-N, anti-LC3B, and anti-β-tubulin (loading control) antibodies. B Cell lesions in CDV-infected Vero and siRNA-ATG5-Vero cell at 32 hpi (magnification, × 4). D-E Vero cells were pre-treated and infected as described in (C). At 24, 28 and 32 hpi, the target protein levels relative to the β-tubulin levels were determined by densitometry. F-G Vero cells were pre-treated and infected as described in (C). At 24, 28 and 32 hpi, copy numbers of CDV were measured by qRT-PCR; virus titers were measured by TCID₅₀. The blots were cropped. The samples derived from the same experiment and that blots were processed in parallel. The data represent the mean ± SD of three independent experiments. Two-way ANOVA; *P < 0.05; **P < 0.01; ***P < 0.001