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. 1999 Nov 29;147(5):1009–1022. doi: 10.1083/jcb.147.5.1009

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© 1999 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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Activation of endogenous Rac by Tiam1 requires membrane localization of the GEF and inhibits migration of NIH3T3 fibroblasts. (A) Activation of endogenous Rac in control NIH3T3 cells or cells expressing C1199- or C580Tiam1. Lysates of the indicated cell lines (from 3 × 10⁶ cells) were incubated with GST-PAK-CD and the bound Rac molecules were detected by Western blot. Aliquots of the respective lysates serve as controls for analyzing total amounts of Rac protein. (B) Tiam1 is a specific activator of Rac and not Cdc42. Lysates of control or Tiam1-expressing NIH3T3 cells were incubated with GST-PAK-CD and the precipitated molecules probed with antibodies against Cdc42. (C) Migration rates of 7 × 10⁴ control or C1199Tiam1-expressing NIH3T3 cells per well coated with fibronectin were determined towards 20 ng/ml PDGF contained in the lower chamber of the transwell. Each bar represents the mean ± SD of triplicate migration assays. Shown is one representative example of three independent experiments.

Activation of endogenous Rac by Tiam1 requires membrane localization of the GEF and inhibits migration of NIH3T3 fibroblasts. (A) Activation of endogenous Rac in control NIH3T3 cells or cells expressing C1199- or C580Tiam1. Lysates of the indicated cell lines (from 3 × 10⁶ cells) were incubated with GST-PAK-CD and the bound Rac molecules were detected by Western blot. Aliquots of the respective lysates serve as controls for analyzing total amounts of Rac protein. (B) Tiam1 is a specific activator of Rac and not Cdc42. Lysates of control or Tiam1-expressing NIH3T3 cells were incubated with GST-PAK-CD and the precipitated molecules probed with antibodies against Cdc42. (C) Migration rates of 7 × 10⁴ control or C1199Tiam1-expressing NIH3T3 cells per well coated with fibronectin were determined towards 20 ng/ml PDGF contained in the lower chamber of the transwell. Each bar represents the mean ± SD of triplicate migration assays. Shown is one representative example of three independent experiments.