Analysis of expression of Lu and B-CAM in HT1080F control cells and transfectants.
A, immunoblot analysis of Lu/B-CAM in control/HT1080F (lane 1), Lu/HT1080F (lane 2), and B-CAM/HT1080F (lane 3) cells. Cell lysates were immunoprecipitated with anti Lu/B-CAM mAb (BRIC221). Immunoprecipitates were separated on a 10% gel under reducing conditions and immunoblotted with anti-Lu/B-CAM polyclonal antiserum. Lu and B-CAM migrated at 80 kDa and 76 kDa, respectively. Molecular weight standards are indicated. B, flow cytometric analyses of Lu/B-CAM and integrin α3/α6/β1/β4 expression. The cells were incubated with the indicated primary antibodies and then with Alexa Fluor 488-conjugated secondary antibody for flow cytometric analysis. The expression of Lu/B-CAM and integrin α3/α6/β1/β4 is shown as a solid line. The gray area indicates the negative control. Overexpression of Lu or B-CAM did not influence the expression of laminin α5-binding integrins α3β1, α6β1, or α6β4.