Figure 3. CSFV infection enhances autophagic flux. (A and B) PK-15 (A) and 3D4/2 (B) cells were mock-infected or infected with CSFV (MOI = 1) for 12, 24, and 48 h. The cell samples were then analyzed by immunoblotting with anti-SQSTM1 and anti-ACTB (loading control) antibodies. (C and D) PK-15 (C) and 3D4/2 (D) cells were infected with CSFV (MOI = 1) in the presence or absence of E64d (10 μg/ml). At 24 and 48 hpi, cell lysates were prepared and analyzed by immunoblotting using anti-LC3B, anti-SQSTM1, anti-E2, and anti-ACTB (for detection of ACTB as loading control) antibodies. The relative levels of the targeted proteins were estimated by densitometric scanning, and the ratios were calculated relative to the ACTB control. The data represent the mean ± SD of 3 independent experiments. Two-way ANOVA; *P < 0.05; **P < 0.01; ***P < 0.001; ^# P > 0.05.