Figure 5.
Integrin suppression requires new integrin ligation. (A) BAE cells plated on Coll I were treated with the α2β1 nonblocking antibody 12F1 or the α2β1-blocking antibody R2–8C8 (20 μg/ml for 60 min). Cells were stimulated with shear stress for 5 min, and GST-FNIII9-11 binding was assessed. Values are means ± SD normalized for total protein (n = 3). **p < 0.01 (B) BAE cells plated on Coll I were treated with 12F1 or R2–8C8, sheared for 5 min, and WOW-1 binding was assessed. Values are means ± SD normalized for total protein (n = 4). *p < 0.05. (C) BAE cells plated on FN were treated with the FN nonblocking antibody 11E5 or the FN-blocking antibody 16G3 (20 μg/ml for 60 min). Cells were stimulated with shear stress for 10 min and binding of biotinylated IAC-1 was assessed by Western blotting with HRP-conjugated streptavidin. Values are means ± SD normalized for total protein (n = 5). **p < 0.01.