Figure 4: Migration and Tube formation assays. (A) Migration assay: Photographs of HUVEC cells from the underside of Boyden chamber membrane. Number of cells migrated in ICM (incomplete medium) used as negative control. Number of migrated cells in ICM with 25 ng/ml bFGF or with bFGF and 1μM tumstatin or 1 μM a1(IV)NC1 were viewed using Olympus CK2 light microscope and representative fields (200x magnification) are shown. (B) Migration assessment assay: The graph displays average number of cells migrated in three independent experiments. (C) Tube formation assay: Tube formation was evaluated after 24 hrs on Matrigel™ matrix with and without 1μM tumstatin or with 1 μM a1(IV)NC1 proteins. Tube formation was visualized in Olympus CK2 light microscope and representative fields at 100x magnification are shown. (D) Tube formation assessment: The graph displays average number of tubes in three independent experiments. In A and C, arrows indicate migrated endothelial cells and tube formation respectively.