Fig. 5.

Autophagy promotes BVDV replication in MDBK cells. (a) Effect of drugs on BVDV-mediated autophagy. MDBK cells were treated with 3-MA or RAP at 0.5 mM and 2.5 μM for 12 h and then infected with HJ-1 or NY-1 at an m.o.i. of 5 or 2.5 for 72 h. The cell samples were then collected for western blot analysis. (b, c) Analysis of the band intensity ratio of LC3-II or E₂ to GAPDH. The results are indicated by graphs representing the ratio of LC3-II or E₂ to GAPDH normalized to the control. (d) BVDV-mediated autophagy in shBCN1-MDBK cells. shBCN1-MDBK cells or control-MDBK cells were infected with HJ-1 or NY-1 at an m.o.i. of 5 or 2.5 for 72 h, and the cell samples were collected for western blot analysis. (e) The viral titres of drug-treated MDBK cells or shBCN1-MDBK cells. After drug-treated MDBK cells or shBCN1-MDBK cells were infected with HJ-1 or NY-1, the cell supernatant of the samples was freeze-thawed three times and used to determine the titer using a TCID₅₀ assay. (f) Relative levels of BVDV RNA. MDBK cells or shBCN1-MDBK cells were treated as described above (a) or (f), and total RNA was isolated. The data are reported as the mean ± SD (n = 3). The data were analysed using a two-tailed Student’s t- test. *, P < 0.05; **, P < 0.01; ***, P < 0.001