A: The association of various Gal-3 proteins with HUVEC at different temperatures. The cells were incubated with 15 µg/ml Gal-3, G13-250, G69-250 or G111-250 for 60 min at 4°C, 20°C, or 37°C. The amount of protein added in the medium is shown in the top panel (medium). Cell associated proteins were analyzed by western blotting with polyclonal antibodies against Gal-3 and actin. B: The exocytosis of various Gal-3 proteins by the cells. The cells were incubated with or without (w/o) Gal-3, G13-250, G69-250 and G111-250 at 4°C for 30 min. After extensive washing, the cells were either lysed immediately (0 min) or incubated in fresh SFM at 37°C for 60 min. After incubation, both the media and the cells were analyzed with western blotting.