14,15-EET activates STAT-3 in HDMVECs. Quiescent HDMVECs were treated with and without 14,15-EET (0.1 μM) for the indicated times and either cell extracts (A) or the cytoplasmic and nuclear factions (C) were prepared and analyzed by Western blotting for pSTAT-3 using its phosphospecific antibodies. (B) The bar graph represents the quantitative analysis of the time course effect of 14,15-EET on tyrosine phosphorylation of STAT-3 in triplicate. Error bars represent SD. The blot in panel A was reprobed with anti–STAT-3 antibodies for normalization. For testing the purity of the cytoplasmic and nuclear preparations, the blot in panel C was reprobed sequentially with anti–STAT-3 and anti-p53 antibodies. *P < .01 versus control.