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Fig. 5. VSMC-dependent regulation of endothelial cell MT1-MMP expression. (A) MT1-MMP expression was monitored in 3D cultures with HUVECs alone or with a combination of HUVECs and VSMCs. With HUVECs alone, MT1-MMP (green) was stained uniformly along the length of growing neovessels (white arrows, left panel). Under these conditions, only low amounts of type IV collagen (red) were detected. By contrast, in HUVEC-VSMC co-cultures, MT1-MMP expression was restricted to the advancing endothelial tip cells (white arrows, right panel) of the capillary stalk positive for type IV collagen, which is comprised of an endothelial cell tube decorated with vascular smooth muscle cells (see supplementary material Fig. S3). Bar, 0.25 mm. (B) In HUVEC-VSMC co-cultures, neovessel stalks express only low levels of MT1-MMP in contrast with strong staining for type IV collagen (panels a-c). By contrast, in the presence of sTek (30 µg/ml), neovessel stalks express heightened levels of MT1-MMP concomitant with low levels of type IV collagen (panels d-f). White arrows indicate areas of MT1-MMP expression. Bar, 0.125 mm.
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