Figure 3

Notch mediates VEGF-induced fibrinolysis and gelatinase activity. A) N1 decoy blocked VEGF-induced fibrinolytic activity. HUVEC were transduced with Ad-VEGF, Ad-N1decoy or both at 40 MOI and Ad-LacZ was used as a control and to normalize the MOI to 80. Transductants were seeded on fibrin gels and fibrin degradation determined after 5 days. B) N1decoy blocked VEGF-induced MMP2 activation. HUVEC were transduced with Ad-VEGF at 20 MOI and increasing amounts of Ad-N1decoy as indicated. Ad-LacZ was used as a control and to normalize the MOI to 80. Four days post-infection, supernatants were collected and subjected to gelatinase zymography. C) N1 decoy blocked VEGF-induced expression and activation of MMP9. HUVEC were transduced with Ad-VEGF at 40 MOI, with or without Ad-N1decoy at 40 MOI and Ad-LacZ was used as a control and to normalize the MOI to 80. Supernatants from transduced HUVEC were collected at 2, 4, 6, and 8 days post-infection and subjected to gelatinase zymography. D) Cross-section of skin from DAS assay stained for PECAM (red) and MMP9 (green). White boxes highlight the large vessels within the subcutaneous adipose depot. Yellow boxes highlight the capillaries within the dermal smooth muscle cell layer. Representative photographs are shown.