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. 2013 Sep 1;24(17):2678–2688. doi: 10.1091/mbc.E13-02-0098

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© 2013 Birukova 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 3: — Effects of Rap1 knockdown on functional and structural barrier restoration of pulmonary EC monolayer after thrombin. (A) ECs plated on microelectrodes were transfected with Rap1-specific siRNA or nonspecific RNA 48 h prior to TER measurements. Control and Rap1-specific siRNA-treated ECs were stimulated with thrombin at the time indicated by the arrow, and TER changes were monitored over time. (B) ECs plated on glass coverslips were transfected with Rap1-specific siRNA or nonspecific RNA prior to stimulation with thrombin. Immunofluorescence staining of F-actin (left) and VE-cadherin (middle) was performed using Texas Red phalloidin and VE-cadherin specific antibody, respectively. Right, merged images of F-actin and VE-cadherin staining. Arrows indicate areas of thrombin-induced intercellular gap formation. (C) Quantitative analysis of gap formation and cell junction VE-cadherin localization in control and Rap1-depleted ECs at different times after thrombin treatment. Data are expressed as mean ± SD; *, p < 0.05.