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Figure 1


Fig. 1. Adhesion to FN causes activation of RhoA and the RhoA-specific GEFs Lsc/p115 RhoGEF and LARG. Mouse fibroblasts were starved in serum-free media and then held in suspension in the same media for 2 hours. (A) Cells were plated onto FN-coated coverslips for the times indicated, then fixed and stained with phalloidin to visualize F-actin, and antibodies against phosphotyrosine to visualize focal adhesions. Bar, 40 µm. (B,C) Cells were plated onto FN-coated dishes for the times indicated, lysed and (B) pulldowns performed with GST-RBD and samples blotted with an antibody against RhoA, or (C) pulldowns performed with GST-RhoA(17A) and samples blotted with antibodies against the indicated GEFs. Quantification of all blots was performed as described in Materials and Methods.





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