Fig. 6

The role of Erk phosphorylation in Lu/BCAM-laminin-related Rac/Rho activity, F-actin arrangement and adhesion of NIH-Lu 11 cells. (A) Phosphorylation of Erk in NIH-Lu11 cells after laminin treatment for different time periods were investigated using Western blotting. (B) The phosphorylation of Erk was evaluated after pretreatment with MEK1/2 inhibitor PD98059 for 1 h at various dosages. (C) F-actin distribution in NIH-Lu11 cells with or without laminin and in the presence or absence of PD98059 was labeled with Alexa Fluor™ 488-conjugated phalloidin. a: Cells without any treatment; b: Cells treated with laminin (0.2 μg/ml); c: Cells treated with laminin (0.2 μg/ml) and PD98059 (50 μM) Arrow: polymeric F-actin. (D) The quantitative data of (C) showed those cells with scattering F-actin distribution. (E) NIH-Lu11 cells were pretreated with PD98059 at various dosages in the presence of laminin. Activities of RhoA and Rac-1 were evaluated by GST-C21 and GST-PAK pull-down assay, respectively, followed by Western blotting. (F) The adhesion ability of NIH-Lu11 cells was evaluated by pretreating cells with PD98059 for 1 h followed by plating cells on 96-well plates pre-coated with laminin 10/11 or BSA. The adhesion ability of NIH-Lu11 cells was then measured. This experiment was repeated three times. *: p < 0.05