Figure 2.

(A) Transient expression of the human b5+b5R protein in COS-7 cells. COS-7 cells (from monkey kidney) were transfected with expression construct (in pCR3.1 vector, Invitrogen) for classical b5R with c-myc tagged at the COOH end or b5+b5R tagged at the NH₂ end. Total cell lysate was collected after 2 days of transient expression and subjected to SDS/PAGE analysis. Approximately 10 μg of total protein was loaded onto each lane and the Western blot was hybridized with mouse antibody 9E10 against c-myc tag. (B) Confocal microscopy for subcellular localization of the human b5+b5R protein. COS-7 cells were grown on glass slide for 2 days after the transfection. Mouse antibody 9E10 and a rhodamine-conjugated donkey antimouse antibody were used to hybridize with c-myc-tagged proteins. b5+b5R (Left) and classical b5R (Right) are shown.