Figure 2.

Detection of West Nile virus (WNV) antigen using western blot in virus-inoculated human corneal epithelial (HCE) cells in vitro. Subconfluent monolayers of HCE cells in 25-cm² flasks were inoculated with WNV at a multiplicity of infection of 0.1 (lanes 1–6) or an equal amount of heat-inactivated virus (lanes 7–12). After 1 hour, the virus inocula were removed, and the cells were rinsed twice in phosphate-buffered saline and incubated in the cell culture medium. Cell lysates were prepared at 1 (lanes 1 and 7), 2 (lanes 2 and 8), 3 (lanes 3 and 9), 4 (lanes 4 and 10), 5 (lanes 5 and 11), and 6 (lanes 6 and 12) days postinoculation. Equal amounts of total protein (8 μg) were resolved on 8–16% Tris-glycine gels and analyzed by western blot using (A) a pooled suspension of anti-WNV E protein monoclonal antibodies or (B) anti-human β-tubulin polyclonal antibody.