FIG 3.

knockdown of autophagic signaling impairs IBDV replication. DF-1 cells were transfected with competent shRNA targeted to Av-LC3B (A), siRNA targeted to Av-ATG5 (B), or scrambled shRNA or scrambled siRNA using Lipofectamine 2000. At 48 h posttransfection, cells were infected with IBDV Gt at an MOI of 0.1. The intracellular and extracellular viral loads were analyzed by WB and TCID₅₀ assay at 24 h p.i. Antibodies against LC3B, β-actin, ATG5, and (p)VP2 were used for WB, and the intensity band ratio of pVP2/actin (C) was normalized to the control. Progeny viral loads both intracellular (D) and extracellular (E) were titrated in a TCID₅₀ assay with secondary CEFs. Means ± SDs were analyzed for three independent experiments. t test was used for comparison to the controls.