Fig. 3

AGE binding activity of galectin-3

Purified recombinant rat galectin-3 (3 µg/dot) was immobilized onto a nitrocellulose filter. After blocking with 2% BSA in blocking buffer for 2 hr filters were probed with increasing concentrations of ¹²⁵I-AGE-BSA (solid line) or ¹²⁵I-BSA (dotted line) in the presence or absence of 100-fold excess unlabeled ligand. After 1 hr, the filters were washed and counted for bound radioactivity. Specific binding was calculated by subtracting nonspecific from total filter-associated radioactivity. (Inset) Scatchard analysis of binding data (representative of three independent experiments).