Effect of deoxygenation on band 3 retention in membrane cytoskeletons isolated from wild-type and transgenic mouse erythrocytes. (A) Erythrocytes from wild-type (left panel), transgenic mice containing a human deoxyHb-binding site (middle panel), or transgenic mice lacking a deoxyHb-binding site (right panel) were either oxygenated or deoxygenated prior to solubilization in 1% Triton X-100. Detergent-insoluble membrane fractions containing the spectrin/actin membrane skeletons were separated by SDS-PAGE and immunoblotted with anti-cdb3 or antiactin antibodies. (B) Densitometric analysis of the relative amount of band 3 retained in the above cytoskeletal pellets using actin as a loading control. All values of band 3 retention in cytoskeletons derived from deoxygenated RBCs are relative to the amount of band 3 retained in the pellet of the corresponding oxygenated erythrocytes (mean of 3 experiments ± standard deviation [SD]).