Fig. 2

CDV infection enhances autophagic flux. A Vero cells were mock-infected or infected with CDV (MOI = 1) for 24, 28, 32, 36 h and 40 h. The expression levels of p62/SQSTM1, CDV-N, and β-tubulin (loading control) were analyzed by western blot with specific antibodies. B The p62 levels relative to the β-tubulin levels were determined by densitometry. C Vero cells pre-treated with rapamycin for 4 h and then mock-infected with DMEM, and cells infected with CDV (MOI = 1) were further cultured in the absence and presence of 10 mg/mL E64d for 24 and 28 h. The cell samples were then analyzed by western blot with anti-CDV-N, anti-LC3B, anti-p62, and anti-β-tubulin (loading control) antibodies. D, E and F The CDV-N, LC3B and p62 levels relative to the β-tubulin levels were determined by densitometry in mock-infected, rapamycin-pre-treated, and CDV-infected Vero cells in the absence and presence of E64d. G Cells were transfected with pmCherry-GFP-LC3B for 24 h, followed by CDV infection (MOI = 1) and treatment with E64d. The fluorescence signals were visualized by confocal immunofluorescence microscopy. H Vero cells were mock-infected or infected with CDV (MOI = 1) for 28 h. The cells were fixed and processed for indirect immunofluorescence using antibodies against LC3 and LAMP1 protein. 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