Fig. 1.

ASFV E199L protein triggers autophagy in Vero and HEK-293T cells. A Vero cells were transfected with empty vectors or various plasmids expressing Flag-tagged ASFV E183L, E199L, F317L, MGF 505-4R, CP530R, B602L proteins and empty vector (EV). At 24 h post-transfection, cells were harvested and Western blotting was performed. Blots are representative of the 3 independent experiments. β-actin was used as sample-loading control. B Densitometric LC3-II/LC3-I and P62/β-actin ratios from at least 3 independent experiments were shown. Error bars show standard error of the mean (SEM). Significance was analyzed with two-tailed Student's test. ∗∗∗P < 0.001. C and E: Vero and HEK-293T cells were transfected with ASFV E199L protein expression plasmids (E199L) or empty vectors (EV). Cells were harvested at indicated time points (6, 12, 24 and 36 h) and detected with anti-LC3B antibody. Blots are representative of the 3 independent experiments. β-actin was used as sample-loading control. D and F: Densitometric LC3-II/LC3-I ratios from at least 3 independent experiments were shown. Error bars show standard error of the mean (SEM). Significance was analyzed with two-tailed Student's test. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001. G and H: Vero and HEK-293T cells were transfected with ASFV E199L protein expression plasmids or empty vectors. The fluorescent puncta of LC3B were observed by confocal microscopy with scale bars indicating 10 μm.