Fig. 4

The effect of Lu/BCAM together with laminin on the activities of RhoA and Rac1 for cell adhesion of NIH-Lu 11 and bladder cancer cells. a Activities of RhoA (upper panel) and Rac1 (lower panel) in NIH3T3 and NIH-Lu 11 cells in the presence of laminin were evaluated by GST pull-down assay. b The effect of Lu/BCAM with or without laminin on the activities of RhoA and Rac1 in the presence of the RhoA inhibitor- C3 exoenzyme. Cells were treated with C3 for 1 h and whole cell lysate was prepared for pull-down by GST-PAK or -C21 protein, followed by Western blotting for activities of RhoA and Rac-1. c Cells were pretreated with C3 of various dosages and plated on 96-well plates, which were pre-coated with laminin 10/11 or BSA. The adhesion ability of NIH-Lu11 cells was then measured and quantitated. These experiments were repeated three times. *: p < 0.05. d The adhesion of five uroepithelial cell lines was further evaluated by cell adhesion assay in the presence of laminin or BSA.*: p < 0.05; **: p < 0.01; ***: p < 0.001. e Activities of RhoA and Rac1 in TCCSUP and J82 cell lines in the presence of laminin were determined by GST-pull down assay followed by Western blotting to illustrate active form or total amount of RhoA and Rac1. The number indicated under each band represents fold difference of active form of RhoA and Rac1 with laminin compared to RhoA and Rac1 without laminin (set as 1). In addition, values were normalized with total RhoA and Rac1 in the cell lysate first