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. 2020 Jun 10;9(11):e016099. doi: 10.1161/JAHA.120.016099

Figure 5. Nck (noncatalytic region of tyrosine kinase) 1 and Nck2 bind to PECAM‐1 (platelet endothelial cell adhesion molecule‐1) in response to shear stress but Nck1 is required for bridging PAK (p21‐activated kinase) 2/PECAM‐1 and activation.

Figure 5

A, Immunoblotting showing PECAM‐1 coimmunoprecipitations and confirms the interactions with Nck1 and Nck2 are shear stress dependent. B, Graphical representation of the levels of Nck1 in PECAM‐1 coimmunopreciptation showing statistical significance. C, Immunofluorescence staining to PECAM‐1 (red) and Nck1/2 (green) after shears stress. Scale bar=50 μm. D, phospho‐PAK2 is detected in PECAM‐1 coimmunopreciptation and (E) PAK membrane translocation and PECAM‐1 binding is enhanced by shear stress and impaired after knocking down of Nck1. F and G, PAK2 activation as assessed by phosphorylation is downregulated by Nck1 ablation but not by Nck2 deletion. Western blots are from n=4 independent experiments. Data are mean±SEM analyzed by either 1‐way ANOVA and Tukey posttest or 2‐way ANOVA and Bonferroni posttest (*P<0.05, **** P<0.0001).