Figure 3.
Gelatinases in human seminal plasma do not cleave galectin-3. A: Eluted fractions from gelatin affinity column chromatography were subjected to SDS-PAGE and silver staining. B: Gelatinase activity was assessed by gelatin zymography. Bacterial collagenase and the soluble fraction of seminal plasma were included as positive controls (upper panel). Immunoblot analysis was performed with anti-MMP-2 and -9 polyclonal antibodies (lower panel). C: The galectin-3 cleavage assay was performed with increasing amounts (by volume) of purified gelatin-binding proteins and of the column flow-through, preincubated with or without 200 mM EDTA. Percent cleavage of galectin-3 was plotted against increasing amounts of flow-through (FT), flow through-EDTA (FT-EDTA), gelatin-binding proteins (GBP) and gelatin-binding protein-EDTA (GBP-EDTA).