Figure 2

TGZ inhibits stress fiber and focal adhesion formation. (A) Effect of TGZ on the distribution of paxillin and stress fibers. ES-2 cells were plated on FN (10 μg/mL)-coated coverslips and incubated in 10% FBS medium with 0.1% DMSO or 20 μM TGZ for 4 h. Cells were then double stained with rhodamine-labeled phalloidin and antibodies to paxillin (FITC). These results are representative of triplicate experiments. (B) Time-course study of the tyrosine phosphorylation level of focal adhesion kinase (FAK) in TGZ-treated ES-2 cells. ES-2 cells were treated with 20 μM TGZ for different intervals. The cell homogenates were immunoprecipitated from 1 mg of total cellular protein by anti-FAK antibody and subjected to Western blot analysis with anti-phosphotyrosine antibody (p-FAK) or anti-FAK antibody (FAK). The cell homogenates were also analyzed for β-actin levels by Western blotting as indicated. Immunoblot results are from a representative experiment performed in triplicate with β-actin as loading control. Symbol (*) indicates cells that were treated with DMSO for 8 h.