Fig 4. In-gel inhibition of MMP-2 and MMP-9 activity by sponge extracts.

MMP-2 and MMP-9 standard were applied to gelatin-zymography. After the electrophoretic run in SDS, gels were cut in lanes and each lane was washed with Triton X-100 and incubated overnight in the incubation medium (50 mMTris; 1% Triton-X-100; 10 mM CaCl₂; 0.2% NaN₃; pH 7.5) in the absence or in the presence of the different aqueous extracts at the highest non-cytotoxic concentrations. 1,10 PA was used as a positive control. A: Staining and destaining of the gels revealed that T. citrina, H. perlevis, I. variabilis and S. spinosulus but not T. aurantium, C. nucula and A. aerophoba, were able to inhibit both MMP-2 and MMP-9. B: Percentage of MMP-9 and MMP-2 inhibition of repeated analyses, calculated in comparison to negative control (CTRL) represented by the lane incubated in the absence of the sponge extracts. Asterisks represent values statistically different from control (One-way Anova followed by Tukey test; * = p<0.05).