Figure 1

Inhibition of ¹²⁵I-VEGF binding to NRP1-expressing carcinoma cells by EG3287 and the effects of VEGF on cell migration. (A) Confluent A549 cells were incubated for 2 h at 4°C with 0.1 nM ¹²⁵I-VEGF in the presence of the indicated concentrations of EG3287. (B) Carcinoma cells were placed into the top inserts and either no addition (control, C), 25 ng ml⁻¹ VEGF, or 0.5 or 1% FBS was added to the bottom wells of the transwell plates. Chemotaxis of these cells towards VEGF or serum was determined after 4 h of incubation. ^*P<0.05; ^**P<0.01 vs control. (C) Vascular endothelial growth factor was measured by ELISA in the conditioned medium collected from cultured A549 cells at the indicated time points after incubation of cells in serum-free medium. Values represent means±s.d. of VEGF (pg ml⁻¹). (D) A549 cells were transfected with control (C) or VEGF siRNA and incubated in serum-free medium for the time periods indicated, after which VEGF was measured in the conditioned medium. (E) A549 cells transfected with control or VEGF siRNA were placed into the top inserts, either no addition (C) or 25 ng ml⁻¹ VEGF was added to the bottom wells of the transwell plates and chemotaxis towards VEGF was determined after 4 h of incubation. ^***P<0.001 vs control siRNA. (F) VEGF siRNA-transfected cells were pretreated for 30 min with EG3287 at 100 μ M and chemotaxis towards 25 ng ml⁻¹ VEGF was determined after 4 h of incubation. ^*P<0.05 vs VEGF siRNA without EG3287.