Fig. 2

2-Ethoxystypandrone (1) inhibited the IL-6-induced and constitutive activation of STAT3. a. Dose-dependent inhibition of 2-ethoxystypandrone (1) on the IL-6-induced STAT3 activity. HepG2/STAT3-luciferase cells were pretreated with 2-ethoxystypandrone (1) at indicated concentrations for 1 h, and the luciferase activity was measured following stimulation of IL-6 (10 ng/mL) for 5.5 h; b. HepG2/STAT3-luciferase cells were treated with indicated concentrations of 2-ethoxystypandrone (1) for 6.5 h. Cell viability was determined by MTT assay; c. Compound 2-Ethoxystypandrone (Cpd 1) inhibited the IL-6-induced phosphorylation of STAT3 in HepG2/STAT3-luciferase cells. HepG2/STAT3 luciferase cells were pre-treated with 2-ethoxystypandrone (1) at the indicated concentrations for 2 h and were then stimulated with 10 ng/mL interleukin-6 for 15 min and the cell lysates were prepared for Western Blot analysis using anti-phospho-STAT3 antibody; d. 2-Ethoxystypandrone (1) inhibited the basal constitutive activation of STAT3 in HepG2 cells. The HepG2 cell lysates were prepared for Westerm Blot after cells were pre-treated with 2-ethoxystypandrone (1) for 2 h and without IL-6 treatment