The anti-angiogenic activity of A009 and HT alone were tested in HUVECs using a morphogenesis assay. HUVECs (5 × 104 cells/well) were plated on a 24-multiwell plate,precoated with 10 mg/mL extracellular membrane basement matrix (Matrigel), then treated for 6 hours with decreasing dilutions (1:500- 1:250) of A009, HT (A) or EtOH (data not shown) in complete M199 medium. Complete medium (CTRL+, M199 supplemented with 10% FBS) and serumfree medium (SFM) were used as positive and negative controls, respectively. The number of meshes were quantified with the ImageJ software “Angiogenesis Analyzer” tool (B). A009 and HT were both able to inhibit morphogenesis in an equivalent manner. Mean ± SD of 3 independent experiments is shown. Student' s t-test was used to determine p-value (*p<0.05)
