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Figure 9


Fig. 9. Inhibition of PKC{alpha} prevents activation of RhoA induced by syndecan-4 clustering. REF cells were plated on FN110-coated dishes for 1.5 hours and DMSO or Gö6976 (1 µM) were added for a further 30 minutes. Subsequently, cells were stimulated with recombinant HepII (1 µg/ml; A), or LPA (400 ng/ml; B). Cells were lysed at the indicated time points and GTP-bound RhoA was captured on GST-Rhotekin-RBD-conjugated glutathione-agarose beads. Samples were resolved by SDS-PAGE, immunoblotted for RhoA and the relative activities of the protein at the indicated time points under each condition determined densitometrically. {blacksquare}, DMSO; {blacktriangleup}, Gö6976. (A) Results were obtained from three independent experiments (mean + s.e.m.; n=3). **P<0.05 compared with levels in Gö6976-treated cells. (B) Results obtained from one of three independent experiments are shown in the graph. The inset shows the ratio of relative RhoA activity between Gö6976- and DMSO-treated cells at various time points following LPA stimulation (mean ± s.e.m.; n=3).





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