FIG. 8.
The furin inhibitor diminished DHBV infection of primary duck hepatocytes. (A) PDH (7 × 105) were incubated with DHBV viremic serum (∼5 × 108 virus particles) overnight, followed by a wash with the medium. Cells were harvested 7 days later (day 7 postinfection [p.i.]). The furin inhibitor (decanoyl-RVKR-chloromethylketone) was added either beginning 12 h prior to incubation with the viremic serum (pre) or immediately following removal of the inoculum (post). DHBV L protein, replicative DNA, and ccc DNA were analyzed. β-Actin served as a control for possible cell toxicity. Lanes a, no inhibitor; lanes b, 20 μM inhibitor; lanes c, 100 μM inhibitor. (B) Southern blot analysis of the ccc DNA at day 2 postinfection. The furin inhibitor was added immediately following withdrawal of the inoculum (same as “post” for panel A). (C) The furin inhibitor was added 2 days after virus infection. Viral replication was detected by Southern blot analysis. The left panel shows the different forms of the viral DNA, including relaxed circular (R.C), linear (L), and single stranded (SS).