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. 2010 Apr 12;207(4):881–896. doi: 10.1084/jem.20091258

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© 2010 Stockton et al.

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 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 3. — Krit1^+/− mouse endothelial cells have increased in vivo and in vitro pMLC and in vitro leak. (A) Endothelial cells were isolated from Krit1^+/+ and Krit1^+/− mouse brain, lung, liver, and kidney as described in Materials and methods, lysed, and Western blotted for immunoprecipitated KRIT1 protein and for pMLC in whole cell lysate. 1, brain; 2, lung; 3, liver; 4, kidney. Endothelial cells from Krit1^+/− mice had similar KRIT1 protein expression in all organs but ∼1/2 the amount seen in Krit1^+/+ mice. Endothelial cells from all organs in Krit1^+/− mice exhibited increased pMLC content in vitro. (B) Isolated pulmonary endothelial cells grown on FN-coated coverslips were also stained for pMLC and f-actin content. Krit1^+/− cells exhibited an increase in both. Bar, 50 µm. (C) In vitro permeability assay of isolated pulmonary endothelial cells. Krit1^+/− endothelial monolayers were twofold more permeable than WT. Permeability was reduced by the ROCK inhibitor H-1152. Error bars are means ± SE (n = 6). *, P < 0.001; **, P < 0.05 compared with vehicle-treated Krit1^+/+.