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. 1999 Sep 6;146(5):1107–1116. doi: 10.1083/jcb.146.5.1107

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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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graphic file with name JCB9906058.f6a.jpg — ERK activity, but not CAS/Crk association, is necessary for cell contraction of collagen gels. Three-dimensional collagen gels containing COS-7 cells were released from the culture dish and allowed to contract for various times as described in Materials and Methods. (A) COS-7 cells in collagen gels pretreated for 60 min with the MEK inhibitor PD98059 (50 μM), the MLCK inhibitor M7 (1 μM), or an inhibitor of myosin ATPase activity (BDM, 10 mM). (B) COS-7 cells transfected with either the empty expression vector (control) or the vector containing CAS without a substrate domain (CAS-SD) or a dominant negative form of MLCK (MLCK−) (Klemke et al. 1997). Contraction is presented as change in diameter (starting-final) measured in millimeters. (C) Cells treated as described in A were washed to remove inhibitors then cultured an additional 24 h before being allowed to contract the collagen matrix for various times as described in Materials and Methods. Each point represents the mean ± SEM of three independent experiments.

graphic file with name JCB9906058.f6b.jpg — ERK activity, but not CAS/Crk association, is necessary for cell contraction of collagen gels. Three-dimensional collagen gels containing COS-7 cells were released from the culture dish and allowed to contract for various times as described in Materials and Methods. (A) COS-7 cells in collagen gels pretreated for 60 min with the MEK inhibitor PD98059 (50 μM), the MLCK inhibitor M7 (1 μM), or an inhibitor of myosin ATPase activity (BDM, 10 mM). (B) COS-7 cells transfected with either the empty expression vector (control) or the vector containing CAS without a substrate domain (CAS-SD) or a dominant negative form of MLCK (MLCK−) (Klemke et al. 1997). Contraction is presented as change in diameter (starting-final) measured in millimeters. (C) Cells treated as described in A were washed to remove inhibitors then cultured an additional 24 h before being allowed to contract the collagen matrix for various times as described in Materials and Methods. Each point represents the mean ± SEM of three independent experiments.