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Fig. 8. Effect of ILK silencing on intracellular signaling pathways. (A, top) Activation of RhoA was determined in control or ILK siRNA-transfected cells after adhesion for the indicated times to fibronectin (FN)-coated plates (10 µg ml–1). Quantification of RhoA activation at 60 minutes is shown below; data represent mean values ± s.e.m. from three independent experiments. (B) Activation of FAK and PAK in cells following adhesion of cells to 10 µg ml–1 fibronectin (FN) for the indicated time was evaluated by western blotting using anti-pFAK (pY397) and anti-phospho-PAK1 antibodies. (C) Translocation of cortactin to membrane ruffles (a Rac-dependent event) was determined in control and ILK siRNA-transfected cells treated with 0.5 µM sphingosine-1-phosphate (S1P) for 5 minutes by immunostaining with monoclonal anti-cortactin antibody. Scale bar, 40 µm.





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