Figure 3

AG490 inhibits constitutive activation of Jak and Stat in HTLV-1-infected T-cell lines and primary ATL cells. (A and B) HTLV-1-infected T-cell lines were treated with increasing concentrations of AG490 for 24 h. (C) Primary ATL cells were treated with (+) or without (-) 50 μM AG490 for 24 h. Phosphorylation status of Stat3 and Stat5 was assessed by Western blot analysis. (D) HTLV-1-infected T-cell lines were treated with increasing concentrations of AG490 for 24 h. Phosphorylation status of Jak1, Jak2 and Jak3 were assessed by Western blot analysis. (E) AG490 does not affect phosphorylation of other phosphor-protein that is not regulated by Jak-Stat pathway. HTLV-1-infected T-cell lines were treated with (+) or without (-) 50 μM AG490 for 24 h. Phosphorylation status of GSK-3β was assessed by Western blot analysis. (F) AG490 inhibits constitutive Stat3- and Stat5-DNA binding in HTLV-1-infected T-cell lines. Nuclear extracts were isolated from HTLV-1-infected T-cell lines treated with (+) or without (-) 50 μM AG490 for 24 h. Stat-DNA binding activity was assessed by EMSA using SIE or β-casein probe.