Effect of aggretin on Matrigel capillary tube formation and angiogenesis in the chick CAM model. (A) HUVECs (2 × 105/well) were treated either in the absence (PBS control, i) or in the presence of VEGF (30 ng/mL, iv); in the presence of aggretin (0.1 μM, ii; or 0.3 μM, iii); or preincubated with anti–integrin α2 mAb, A2-IIE10, for 30 minutes, then treated in presence of aggretin (0.1 μM, v), and then plated on diluted Matrigel (4 mg/mL) for 24 hours. After washing and fixation, cells were observed under microscope at × 40 magnification and photographed. A representative one of 4 similar experiments is shown. (B) CAMs of 10-day-old chick embryos were incubated with vehicle (PBS, i), VEGF (200 ng/disk, ii), aggretin (0.6 μg/disk, iii), or compounds in combination with A2-IIE10 (50 μg/mL, iv) or VEGF mAb (1 μg/mL, v-vi) for 48 hours, and then resected, fixed, and photographed with a stereomicroscope at × 10 magnification. A representative one of 3 similar results is shown.
