Figure 5.

bFGF and VEGF₁₆₅ interfere with LfcinB binding to immobilized heparin. Biotinylated LfcinB (50 μg/ml) alone or in combination with the indicated concentrations of bFGF, VEGF₁₆₅, or nonheparin-binding EGF was added to heparin-coated wells of a flat-bottom tissue culture plate and incubated for 2 hours. Plates were then washed, and streptavidin-HRP was added to wells. After 2 hours, plates were washed and 1,2-phenylenediamine substrate was added. Absorbance was measured at 492 nm. Data are shown as mean absorbance ± SD of quadruplicate determinations. Background absorbance was 0.043 ± 0.001. Statistical significance relative to the LfcinB control was determined by the Tukey-Kramer multiple comparisons test; *P < 0.001.