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

Figure 6

Resveratrol protected HUVEC from ROS-induced loss of adhesion and from FAK/ERK dephosphorylation. Cells were preincubated with 5 and 10 μM of Resveratrol for 4 hours and then treated with Hx-XO (4.5 mU/mL) for 60 minutes. (a) Adherent HUVECs were determined by loading cells with 2.5 μM Calcein-AM as described in Materials and Methods. (b) The cells were harvested and the soluble fraction was analysed for FAK activation. FAK was immunoprecipitated using a monoclonal antibody, and its phosphorylation state was analysed by Western blot with anti-phosphotyrosine. (c) Cells were harvested 30 minutes later for analysis of ERK1/2 activity by Western blot with anti-phospho-ERK1/2 antibodies. Results are expressed as mean ± SD of three experiments, each performed in triplicate. compared to the Hx-XO alone.

(a)

(b)

(c)