Peripheral blood smear, red cell membrane proteins, and glycophorins before transfusion.

(A) Red blood cells with bizarre shapes, spherocytes, and erythroblasts with cytoplasmic elongations. (B) Coomassie blue staining. (C) Western blotting of band 3 using a monoclonal antibody (Sigma, St Louis, MO). (D) Western blotting of protein 4.2 using our own polyclonal antibody. (E) Silver staining of glycophorins; a: control; b: patient (venous blood prior to transfusion). (B) In the patient, band 3 and protein 4.2 were missing (denoted by ←); Spectrin α and β chains, ankyrin were strongly reduced, as was band 6 (*). Unlike the membrane of mice with targeted inactivation of the band 3 gene,9 10 the patient's membrane retained negligible amounts (4%) of hemoglobin (versus 1% in controls); residual hemoglobin would not significantly alter the quantification of membrane proteins. The amounts of protein 4.1 and actin were found to be rather similar to those of controls. (C) Band 3 was present and accompanied by known proteolytic fragments (60, 41, and 22 kd) in the control. In the patient, band 3 was totally missing (←). (D) Protein 4.2 was totally missing as well (←). (E) There was a strong reduction of the GPA dimer (**) and a less pronounced decrease of the GPA monomer (*). The heterodimer GPA/GPB was nearly undetectable (←). No obvious differences were recorded as for the other glycophorins.