Figure 3

Effect of modulators of angiogenesis on the activity and production of MMPs: Equivalent aliquots of the medium normalized to the total cell protein from the culture of control, ursolic acid (50 μM), curcumin (10 μM) and aspirin (100 μM) treated cells on days 1 to 4 were subjected to zymography (A) as described in the text.(a) Lanes 1-4, 50 μM ursolic acid treated cells 1^st -4^th day, Lanes 5-8, control 1^st-4^th day. (b) Lanes 1-4, 10 μM curcumin treated cells 1^st -4^th day, Lanes 5-8, aspirin treated cells 1^st-4^th day. The activity of MMP-9 and MMP-2 in zymogram was quantitated using Quantity One 4.5.0 software BioRad geldoc and expressed in arbitrary units of intensity/mm²/mg protein (B). Values given are the average of intensity measurement of 6 experiments ± SEM. The culture medium collected from control, ursolic acid (50 μM), curcumin (10 μM) and aspirin (100 μM) treated cells at intervals of 24 hrs (normalized to the total cell protein) were quantitated for MMP-9 and MMP-2 protein expression by ELISA using specific antibody (C). Values given are the average of duplicate analysis of 5 experiments ± SEM. * Statistically significant compared to the production on first day (p < 0.05) # Statistically significant compared to controls (p < 0.05).