STAT-5B binds to IL-8 promoter in response to 15(S)-HETE in Jak2-dependent manner. (A,C) Quiescent HRMVECs were treated with and without 0.1 μM 15(S)-HETE for the indicated time periods, and either nuclear extracts were prepared and analyzed by EMSA for STAT-DNA–binding activity using [³²P]-labeled STAT-binding sequence of IL-8 promoter as a probe in vitro (A) or processed for ChIP analysis of STAT-5B binding to IL-8 promoter in vivo (C). (B,D) HRMVECs that were transduced with Ad-GFP, Ad-dnJak2, or Ad-dnSTAT-5B with 80 MOI and quiesced were treated with and without 0.1 μM 15(S)-HETE for 2 hours, and either nuclear extracts were prepared and analyzed by EMSA for STAT-DNA–binding activity (B), as described in (A), or processed for ChIP analysis of STAT-5B binding to IL-8 promoter in vivo (D).