Fig. 4.

ANE induces dephosphoylation of STAT3. OC2 cells cultured with various concentrations of or 1% FBS were treated with 0.5 mg/ml ANE. Twenty-four hours after treatment, cells were directly harvested for Western blot (A) or fractionated for evaluating the localization of STAT3 (B). The effect of ANE on STAT3 phosphorylation in two independent experiments was demonstrated after normalization to each untreated sample (A, lower panel). The value of each untreated sample under different FBS conditions was defined as 1. (C) Cells transfected with IL8 promoter reporter were treated with ANE in combination with NSC74859 or JAK inhibitor I. The used doses of NSC74859 and JAK I are 50 and 1 μM, respectively. The luciferase activity was determined 24 h later.