Figure 5.

Autophagy inhibition reduced the replication of BTV1. (A,E) Effects of 10 mM 3-MA or 20 μM CQ treatment on LC3-II accumulation and VP2 or NS3 expression. The expression of relative proteins were analyzed by immunoblotting with specific antibodies; (B,F) Representative results are shown with graphs representing the intensity of LC3-II/actin; (C,G) Representative results are shown with graphs representing the intensity of VP2/actin. (D,H) Effects of 10 mM 3-MA or 20 μM CQ on the production of infectious particles of BTV1. The extracellular and intracellular virus loads were tested as TCID₅₀/mL at 24 hpi and 36 hpi, respectively; (I) The effects of Beclin1 silencing on LC3-II and VP2 or NS3 expression. BSR cells were transfected with either specific siRNA targeting Beclin1 or negative control siRNA. At 24 h after transfection, the cells were incubated with BTV1 (MOI = 1). Samples were collected 24 hpi and analyzed by Western blotting with antibodies against Beclin1, LC3, VP2, and NS3; (J) Representative results are shown with graphs representing the intensity of VP2/actin; (K) The viral titers were determined as TCID₅₀/mL at 24 hpi. Data represent the mean ± SD of three independent experiments. * p < 0.05 and ** p < 0.01, significantly different.