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. 2001 Mar 19;152(6):1145–1158. doi: 10.1083/jcb.152.6.1145

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© 2001 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Effect of uPAR expression on Rac activity in growing NIH 3T3 cells. (A) NIH 3T3 cells were transfected by electroporation with pRC/CMV-uPAR and/or pRK5–myc-wtRac as indicated. PRK5-myc transfection was used as a control. The levels of total cellular Rac and GST–PAK–CRIB precipitated active Rac were determined by SDS-PAGE on 15% gels followed by immunoblotting for Rac. Results shown for endogenous Rac activation are for cells lysed 4 h after transfection. Results shown for cotransfected wtRac activation are for cells lysed 16 h after transfection. (B) The level of activated Rac from cells lysed 16 h after transfection with pRK5–myc-wtRac or pRK5–myc-wtRac and pRC/CMV-uPAR was quantified by density scanning using NIH Image software. The ratio of activated to total Rac was determined and results are expressed relative to the Rac activity in cells transfected with pRK5–myc-wtRac. Results are mean ± SD from three independent experiments.