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Fig. 6. Quantitation of anti-VEGFR-3 and anti-VEGFR-2 antibody-mediated inhibition of in vitro angiogenesis in BAE cells. Confluent BAE cell monolayers on three-dimensional collagen gels were treated with VEGF-C_NC (100 ng/ml) (A) or VEGF₁₆₅ (100 ng/ml) (B). Anti-human VEGFR-3 antibody (hF4-3C5), anti-human VEGFR-2 monoclonal antibody (IMC-1C11) and control antibodies (anti-KLH for hF4-3C5, IMC-C225 for IMC-1C11) were added. Invasion was measured after 4 days and is expressed as percent of sprouting induced by cytokine alone. (A) hF4-3C5 and IMC-1C11 inhibited VEGF-C_NC-induced BAE cell invasion by 68% at 5 µg/ml and 66% at 20 µg/ml, respectively. A complete inhibition of VEGF-C_NC-induced BAE cell invasion was obtained by the co-addition of both antibodies, whereas the control antibody (KLH) had no effect on invasion at 25 µg/ml. (B) When added to VEGF-A-treated BAE cells, hF4-3C5 antibody had no effect at 5 µg/ml, whereas IMC-1C11 at 20 µg/ml totally blocked invasion. The isotope control antibody for IMC-C225 had no effect on invasion. Results are shown as the means±s.e.m. from at least three experiments per condition.

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