Effects of β3 missense mutations on the expression of αvβ3.
(A) Flow cytometric analysis of αvβ3 on the transfected cell surface. Wild-type β3 cDNA or each mutant β3 was cotransfected into 293 cells with wild-type αv. The binding of LM609 or LM142 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 αvβ3. 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.
