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. 2011 Jul 15;22(14):2509–2519. doi: 10.1091/mbc.E11-02-0157

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© 2011 Liu et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 4: — KRIT1–Rap1 binding regulates HuVEC permeability. (A) Knockdown of KRIT1 expression by KRIT1 siRNA causes an approximately twofold increase in permeability (left). Reconstitution of recombinant WT KRIT1 reverses increased permeability (center). Recombinant KRIT1(R452E) mutant does not reverse increased permeability (right). Data shown are mean increased percentage over control siRNA in permeability ± SEM; n = 3. *p < 0,05 compared with control siRNA. (B) A representative blot of KRIT1 depletion and reexpression of recombinant proteins. More than 80% knockdown of KRIT1 is observed by immunoprecipitation and blotting. Reconstituted wild-type and RE (arginine 452 to glutamic acid) mutant KRIT1 proteins are expressed at equivalent levels. (C, D) Knockdown of KRIT1 expression causes an approximately threefold increase in pMLC/total MCL ratio (left). Reconstitution of recombinant WT KRIT1 reverses MLC phosphorylation increase (center). Recombinant KRIT1(R452E) mutant does not reverse increased pMLC level (right). Data shown are mean increased percentage over control siRNA; n = 2. (D) Immunoblots of pMLC (top) and total MLC (bottom) in HuVECs.