Effect of o-vanadate on the ability of BS3 to cross-link band 3 into high–molecular weight aggregates. Erythrocytes were treated in the presence (lanes 1-5) or absence (lanes 6-10) of 2mM o-vanadate, and then exposed to increasing concentrations of BS3 (0.5-5mM) before preparation of ghosts and analysis of the molecular weights of band 3 species present by polyacrylamide gel electrophoresis followed by Western blotting (A). The bands in panel A were quantified by densitometry using a laser infrared fluorescence detector (Odyssey; LI-COR Biosciences) with Odyssey V3.0 software and plotted against BS3 concentration (B). Lanes 1-5 (without o-vanadate) are plotted in the top curve (CTRL + BS3) and lanes 6-10 (with o-vanadate) are plotted in the bottom curve (o-v + BS3). Band density is expressed as arbitrary fluorescence units and values are means of 3 separate experiments. Bars represent SD from the mean.
