Band 3 and adducin deficiency across sph alleles. (A) Coomassie blue-stained SDS-PAGE gel assessment of RBC ghosts proteins from several sph alleles. Equal volumes of normal and mutant packed RBC ghosts/lane were loaded. (B) Western blots of RBC ghosts proteins probed with the indicated antibodies. Each panel is representative of results from multiple blots. (C) Eosin-5-malemide staining of erythroid band 3 was measured for mature RBCs and reticulocyte gated populations and is presented as MFI. Eosin-5-malemide MFI is significantly (*) reduced (P < .05) in RBCs and reticulocytes of all sph alleles examined except sph4J. (D) Alexa Flour 647 fluorescent intensity staining of erythroid band 3 was measured for combined RBC and reticulocyte-gated populations because no difference was observed between the cell types (data not shown) and is presented as MFI. Alexa Flour 647 MFI is significantly (*) reduced (P < .05) in all sph alleles examined. All plotted values in panels C and D are expressed as mean ± standard error. Values in parentheses below the x-axis represent the percentage of respective control.