Figure 4.

Specificity of bigelovin on JAK2/STAT3 signaling. (A) HeLa cells were treated with bigelovin at the indicated concentrations for 1 h. Then, the cells were lysed for Western blot analysis and probed with anti-tyro-phosphorylation antibody. (B) Effects of 50 μmol/L bigelovin on activities of human kinases. In vitro kinase assays were processed by Millipore Kinase Services. (C) HeLa cells were pretreated with bigelovin at the indicated concentrations for 1 h and then stimulated with EGF (100 ng/mL) or Insulin (100 nmol/L) for 10 min. Cells were harvested for Western blot analysis using the indicated antibodies. (D) HeLa cells were pretreated with bigelovin for 1 h before stimulation with IFN-γ (20 ng/mL) for 15 min. Whole-cell lysates were processed for Western blot analysis with the indicated antibodies. An anti-α-tubulin antibody was used as a loading control.