Figure 1.

Determination of chimeric α5-integrin expression in transfected CHO B2 cells. (A) The X5C5, X5C4, X5C2, X5C0, and P3 cell lines were trypsinized from near confluent plates, washed and incubated with a mAb (mab) specific for the α5 extracellular domain, followed by incubation with a FITC-conjugated secondary antibody as described. The control panel (left) represents cells incubated with secondary antibody only. (B) Attached cells were lysed and total cellular lysates were analyzed by immunoblotting using a mab specific for the cytoplasmic (C) domain of the α5 or the α2 integrin. Alternately, lysates were incubated with a mab specific for the extracellular (X) domain of the α5 subunit. Immunocomplexes were recovered and analyzed by immunoblotting with a mab specific for the cytoplasmic domain of α4 or of α5, as indicated. In both cases, total lysates were analyzed by immunoblotting with a mab specific for the extracellular domain of the α5-Integrin to ensure equal loading.