FIG. 4.
CELISA binding of gHt.st/gL to cells. (A to F) gHt.st/gL, gDΔ290-299 (gD290t), gB730t, or GFPst was allowed to react with cells grown in 96-well plates. Binding was detected by means of appropriate antibodies (PAb R45 to gD, MAb H1817 to gB), followed by HRP-conjugated secondary antibodies or HRP-conjugated MAb to the One-STrEP tag (Strep-Tactin) for gHt.st/gL or GFPst and the o-phenylenediamine substrate. (G to J) gHt.st/gL or GFPst was reacted with the indicated cells in medium adjusted to pH 7, pH 6, or pH 5. (K) CELISA with K562, K562αVβ3, and 293T cells in suspension. gHt.st/gL, gB730t, or GFPst was reacted with cells as described above, except that the cells were in suspension. Binding was detected as in panels A to F. (L to N) Binding of gHt.st/gL to receptor-negative CHO and J cells or to heparan sulfate- and chondroitin sulfate-negative sog9 cells. Abscissa, μM concentrations.