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Figure S5. Dominant-negative FIP2 and Rip11 do not inhibit A2780 cell migration into FN-containing matrigel. (A) Migration of A2780 cells expressing FIP2wt or Rip11wt or their corresponding dominant-negative mutants, FIP2480E and Rip11630E, into matrigel plugs in the presence of 25 µg/ml fibronectin (FN) was determined. 1 µM cilengitide (CIL) was added as indicated to the matrigel and to both the upper and lower chambers of the inverted assay. Migrating cells were stained with Calcein AM and visualized by confocal microscopy. Serial optical sections were captured at 15-µm intervals, and cell migration was quantitated by measuring the fluorescence intensity of cells penetrating the matrigel to depths of 45 µm and greater, and expressing this as a percentage of the total fluorescence intensity of all cells within the plug. Data represents mean ± SEM from three independent experiments. (B) Influence of cilengitide and dominant-negative RCP on surface expression of α5β1 and EGFR1. A2780 cells were transfected with RCPwt or dominant-negative RCP621E. Cells were serum starved and treated with 1 µM cilengitide for 20 min. The quantity of α5β1 and EGFR1 expressed at the plasma membrane was determined by cell surface biotinylation followed by detection of the labeled receptors by capture ELISA. Values are expressed as a percentage of the amount of surface receptors expressed by RCPwt-expressing cells. Values are mean ± SEM; n = 3 independent experiments.