Integrin β3 is dispensable for RhoA activity and contractility.

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Integrin β3 is dispensable for RhoA activity and contractility. Integrin β3 is dispensable for RhoA activity and contractility. (A) Schematic representation of previous results. Itgb1-null cells (GE11) have very low levels of GTP-bound RhoA and fibronectin matrix assembly. Re-expression of β1 integrin, but not overexpression of β3 integrin induces highly dynamic cell-matrix adhesions, a mesenchymal cell morphology and restores RhoA activity and fibronectin matrix assembly. Images show GEβ1 and GEβ3 cells stained for paxillin (green) and F-actin (red). (B) Western blot analysis of RhoA activity assay on lysates of Itgb3 knockout and wild-type MEFs overexpressing indicated integrins. Quantification shows relative RhoA activation ± s.d. compared with wild-type MEFs from two independent experiments. (C) Images of assembled fibronectin-biotin on Itgb3-knockout and wild-type MEFs; fibronectin is red and nucleus blue in merged images. Scale bar: 50 μm. (D) Itgb3-knockout and wild-type MEFs overexpressing indicated integrins stained for paxillin (green), F-actin (red). Nuclei are stained blue with Topro-3. Arrows indicate long actin fibers. Scale bar: 10 μm. (E) Flow cytometry analysis of integrin β1 surface expression on Itgb3 knockout and wild-type MEFs transfected with integrin β1 or control siRNA. (F) Wild-type and Itgb3-knockout MEFs transfected with integrin β1 or control siRNA stained for paxillin (green) and F-actin (red). Two representative examples of integrin β1 siRNA transfected wild-type MEFs are shown. Stephan Huveneers et al. J Cell Sci 2008;121:2452-2462 © The Company of Biologists Limited 2008