Effects of β3 missense mutations on the expression of αIIbβ3.
(A) Flow cytometric analysis of αIIbβ3on the transfected cell surface. Wild-type β3 cDNA or each mutant β3 was cotransfected into 293 cells with wild-type αIIb cDNA. The binding of AP2 or TP80 to the transfected cells was analyzed by flow cytometry 2 days after transfection. Results were mean ± SD from 3 separate experiments and expressed as percent MFI relative to that of wild-type αIIbβ3. Two-tailed P values for paired samples were obtained by the Student t test (*P < .01, **P < .05). (B) Immunoprecipitation analysis of biotin surface-labeled transfected cells. The transfected cells were surface-labeled with biotin 2 days after transfection. Immunoprecipitation was then performed using AP3. Precipitates were separated by 6% SDS-PAGE under reducing conditions. After transfer to a nitrocellulose membrane, precipitated proteins were detected by chemiluminescence.
