FIG. 4.

(A) FACS to detect HSV binding to inducible B5-Tet-ON porcine cells or human HEp-2 cells. Inducible B5-expressing porcine cells were grown in the presence of 1 mg/ml DOX before performing binding assays. (B) Induced B5 cells were exposed to the indicated peptides under different conditions. Cells were pretreated with B5 peptides (wild type [wt] and mutants), and unbound peptide was washed out with PBS before infection with HSV-1(F) (peptide preincubated with cells, left panels). In parallel experiments, purified HSV-1(F) was preincubated with 42 mM peptide, and the mixture used to infect B5-expressing cells. One duplicate sample was washed with heparin to detect stable binding (virus preincubation, middle panels) or with PBS to determine total binding (virus preincubation, right panels). Bound HSV was detected by FACS using antibodies to HSV glycoproteins. In panel A: gray line with an arrowhead, stable bound virus; black line, total virus bound. In panel B: gray line with an arrowhead, stably bound virus in the presence of peptide; black line, total virus bound. The filled peaks indicate mock-treated cells.