Xanthine Oxidase-Derived ROS Display a Biphasic Effect on Endothelial Cells Adhesion and FAK Phosphorylation

Figure 4

Effect of wortmannin, a PI-3 kinase inhibitor on the early phase of FAK phosphorylation and HUVECs adhesion. HUVECs were preincubated for 20 minutes with PI-3 kinase inhibitor wortmannin (100 nM) and then treated with Hx-XO (4.5 mU/mL) for 15 minutes. (a) FAK was immunoprecipitated using a monoclonal antibody and its phosphorylation state and expression were analysed by Western blot with anti-phosphotyrosine and anti-FAK antibodies. (b) Phosphorylated FAK from different experiments was quantified by densitometry and corrected for the amount of FAK which was quantified by densitometry. (c) HUVECs adhesion was measured after 15 minutes of treatment with Hx-XO by loading cells with 2.5 μM Calcein-AM. Results are expressed as % of control without Hx-XO (mean ± SEM, , Hx-XO versus untreated controls; ^#wortmannin versus Hx-XO alone).

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(b)

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