Figure 10. Inhibition of autophagy with LC3B-targeting shRNA reduces the replication of CSFV. (A and E) PK-15 (A) and 3D4/2 (E) cells were transfected with shRNAs targeting LC3B or scrambled shRNAs for 48 h, followed by mock infection and CSFV infection at an MOI of 0.5. At 24 hpi, the silencing efficiency of LC3B shRNA, as well as the expression of autophagy marker proteins and CSFV-E2 were analyzed as described in the legend to Figure 2A. The data represent the mean ± SD of 3 independent experiments. Two-way ANOVA; *P < 0.05; ***P < 0.001. (B and F) PK-15 (B) and 3D4/2 (F) cells were transfected with LC3B shRNA for 48 h, followed by CSFV infection at an MOI of 0.5. At 24 hpi, the cells were analyzed as described in the legend to Figure 8B and F. Scale bar: 10 μm. One of 3 independent experiments is shown. (C and G) PK-15 (C) and 3D4/2 (G) cells were transfected as described in (A and E), followed by CSFV infection at an MOI of 0.5 for 24 h. Both the extracellular and intracellular copy numbers of CSFV were detected by qRT-PCR. The data represent the mean ± SD of 3 independent experiments. Two-way ANOVA; **P < 0.01; ***P < 0.001. (D and H) PK-15 (D) and 3D4/2 (H) cells were transfected and infected as described in (C and G). At 24 hpi, both the extracellular and intracellular virus titers were measured by endpoint dilution titrations by using the immunofluorescence assay described in Materials and Methods. Results are expressed in units of TCID₅₀/ml. The data represent the mean ± SD of 3 independent experiments. Two-way ANOVA; . *P < 0.05; **P < 0.01; ***P < 0.001.