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. Author manuscript; available in PMC: 2011 Jul 1.
Published in final edited form as: Microvasc Res. 2010 Jan 4;80(1):75–88. doi: 10.1016/j.mvr.2009.12.010

Figure 4. Colocalization of EGFP-nmMLCK2 and endogenous cortactin in human lung endothelial lamellipodia.

Figure 4

Panel A. EC overexpressing EGFP-nmMLCK2 were incubated with either vehicle (i), thrombin for 10 min (ii), S1P for 30 min (iii), or thrombin for 10 min followed by S1P for 30 min (iv) at 37°C, then fixed and immunostained for cortactin (red). Yellow indicates areas of colocalization. The lower panels, i′-iv′, depict colocalization of endogenous cortactin with nmMLCK2 as described for Figure 3 above. Shown are representative images selected from 7–13 images for each treatment. Panel B: The mean ICQ values for cortactin and EGFP-nmMLCK2 colocalization were calculated as described in Methods for the entire cell, the cytoplasm, and the lamellipodia under each condition [vehicle (No stim), thrombin for 10 min, S1P for 30 min, or thrombin for 10 min followed by S1P for 30 min]. Thrombin induces a statistically significant reduction in peripheral as well as whole-cell colocalization of nmMLCK2 with cortactin when compared to unstimulated (whole cell or lamellipodia, *p <0.02) and S1P-stimulated cells (whole cell or lamellipodia, *p <0.01). N = 7–13 per condition.