Fig. 5.

Intracellularly-expressed Nb1 inhibits BVDV replication. MDBK, MDBK-Nb35^EGFP and MDBK-Nb1^EGFP cells were infected with 0.05 MOI BVDV for 24, 36, and 48 h, and culture supernatants and cells were collected. Expression in cell lysates was detected by RT-qPCR (A), and western blotting (B) using an anti-NS5B polyclonal antibody or anti-α-tubulin antibody. Extracellular BVDV RNA was analyzed (C) and the virus titers in the culture supernatants were measured based on the TCID₅₀ value (D). MDBK, MDBK-Nb35^EGFP, and MDBK-Nb1^EGFP cells were infected with 1 MOI BVDV for 1, 2, 4, 5, 6, 8, 10, and 12 h, and cells were collected for RT-qPCR (E). Data are shown as the mean ± SD representative of three independent experiments. p values were calculated using ANOVA. Significant differences, as compared to control values are denoted as ***(p < 0.001).