Figure 1

Autophagy is induced in ORFV-infected OFTu cells. A OFTu cells were mock infected or infected with ORFV-CL18 (MOI = 10), and 36, 48 and 60 hpi, the expression levels of the autophagy marker proteins LC3B and P62 were measured by Western blotting. GAPDH was used as the loading control. B, C The ratios of LC3-II to LC3-I B and P62 to GAPDH C were quantified by ImageJ software. The results were expressed as the mean ± SD (n = 3) (**P < 0.01, ***P < 0.001, ****P < 0.0001; unpaired t test). D OFTu cells were mock infected, infected with ORFV-CL18, treated with 30 μM rapamycin (Rapa) or treated with 30 μM Rapa, infected with ORFV and then cultured for 60 h. The expression levels of LC3B and P62 were measured by Western blotting. GAPDH was used as the loading control. E The ratios of LC3-II to LC3-I and P62 to GAPDH after different treatments. The results are expressed as the mean ± SD (n = 3) (*P < 0.05, *** P < 0.001; one-way ANOVA). F OFTu cells were mock infected, infected with ORFV-CL18, treated with 5 mM 3-methyladenine (3-MA) or treated with 5 mM 3-MA, infected with ORFV and cultured for 60 h. The expression levels of LC3B and P62 were measured by Western blotting. GAPDH was used as the loading control. G The ratios of LC3-II to LC3-I and P62 to GAPDH after different treatments. The results are expressed as the mean ± SD (n = 3) (**P < 0.01, *** P < 0.001, **** P < 0.0001; one-way ANOVA). H OFTu cells were mock infected, infected with ORFV-CL18 or treated with 30 μM Rapa after transfection with the GFP-LC3 vector and cultured for 24 h. Autophagosomes were observed by confocal laser scanning microscopy. Scale bar, 10 μm. I Fluorescence images were analysed with ImageJ software, and GFP-positive puncta were quantified, and the results are expressed as the mean ± SD (n = 3) (***P < 0.001; unpaired t test).