Figure 5

Effects of inhibiting STAT5 on TF/FVIIa-induced Bcl-2 expression. A, TF-transfected SK-N-SH cells were incubated with or without 10 nM FVIIa, and elevating doses of JAK inhibitor 1 (100, 300, 600 nM) and LY294002 (5, 10, 20 μM) for 2 h. Bcl-2 expression was detected by western blot assay. B, TF-transfected SK-N-SH cells were treated with different concentrations of either STAT5 siRNA or control siRNA for 24 h. The expression of endogenous STAT5 was detected by western blot analysis. C, TF-transfected SK-N-SH cells were incubated with 10 nM FVIIa in the absence or presence of varying doses of either STAT5 siRNA or control siRNA. Bcl-2 expression was detected by western blot, and densitometric data for the Bcl-2 bands provides the degree of inhibition of TF/FVII-induced Bcl-2 expression achieved by STAT5 siRNA treatment. The western blots shown in Figs A and B are representative of at least three independent experiments, and data shown in Fig C are mean fold inhibition (± SD) of Bcl-2 of three independent assays.