Fig. 5

Inhibition of ERK phosphorylation and immunophenotyping for EC markers. I Concentration -dependent effect of ERK inhibitor (U0126). Three different concentrations (0.5, 1.0, and 5.0 μM) of U0126 were used. Western blot analysis showed significant inhibition of p-ERK by 1.0 and 5.0 μM of U0126. However, 5.0 μM of U0126 showed the highest inhibition among all three different concentrations. Phospho-ERK was normalized to its total protein expression. II Flow cytometric analysis of PECAM1 (CD31) with ERK inhibitor (U0126). Three different groups treated with 5.0 μM of U0126: AMSCs with EGM (A), AMSCs with EGM and MMP-2 siRNA (B), and AMSCs with EGM and MMP-14 siRNA (C). Flow cytometry data were analyzed to show the significant differences between the groups (D). III Flow cytometric analysis of VE-cadherin (CD144) with ERK inhibitor (U0126). Three different groups were treated with 5.0 μM of U0126: AMSCs with EGM (A), AMSCs with EGM and MMP-2 siRNA (B), and AMSCs with EGM and MMP-14 siRNA (C). Flow cytometry data were analyzed to show the significant differences with or without U0126 (D). *p < 0.05, **p < 0.01, ***p < 0.001. EBM endothelial cell basal medium, EGM endothelial cell growth medium, MMP matrix metalloproteinase, ERK extracellular signal-regulated kinase, HUVEC human umbilical vein endothelial cell