Fig. 2
IL-6 induces STAT3 (Y705) phosphorylation. a IL-6 induces STAT3 (Y705) phosphorylation in a dose-dependent manner. SK-Hep-1 cells were treated with the indicated doses of IL-6 for 4 h and analyzed by western blotting using the indicated antibodies. pSTAT3, Y705-phosphorylated STAT3. b Time-course studies of IL-6-induced STAT3 (Y705) phosphorylation. SK-Hep-1 cells were treated with 50 ng/ml IL-6 for the indicated times. Total cellular proteins were prepared for western blotting at the end of each treatment. c STAT3 inhibition blocked IL-6-induced STAT3 activation. SK-Hep-1 cells were cultured in the presence or absence of 5 μM LLL12, a STAT3 inhibitor for 2 h and then with 50 ng/ml IL-6 for an additional 12 h. Total cellular proteins were prepared for western blotting analysis. d STAT3 inhibition blocked IL-6-induced lncTCF7 upregulation. qRT-PCR analysis of lncTCF7 in SK-Hep-1 cells cultured in the presence or absence of 5 μM LLL12, a STAT3 inhibitor for 2 h and then with 50 ng/ml IL-6 for an additional 12 h. e SK-Hep-1 cells were transfected with a negative control, 50 nM control siRNA (siCon) or 50 nM STAT3 siRNA (siSTAT3) as described in the Materials and methods. Forty-eight hours after transfection, the cells were treated with 50 ng/ml IL-6 for 12 h. At the end of the cell culture period, cell lysates were prepared for determination STAT3 activation by western blotting. Data are representative of at least three independent experiments. f The cells were treated as described in section E. At the end of the cell culture period, cells were prepared for qRT-PCR analysis of lncTCF7 (n = 3)