Egr-1 binds to FGF-2 promoter in response to 15(S)-HETE both in vitro and in vivo in Src-dependent manner. (A,C) Quiescent HDMVECs were treated with and without 15(S)-HETE (0.1μM) for the indicated time periods, and either nuclear extracts were prepared and analyzed by EMSA for DNA binding activity in vitro with the use of [³²P]-labeled double stranded oligonucleotide probe that corresponds to the Egr-1 binding site proximal to the transcriptional start site in the FGF-2 promoter (A) or processed for ChIP analysis of Egr-1 binding to FGF-2 promoter in vivo (C). (B,D) HDMVECs that were transduced with Ad-GFP, Ad-dnSrc, or Ad-dnEgr-1 with a moi of 40 and quiesced were treated with and without 15(S)-HETE (0.1μM) for 30 minutes and either nuclear extracts were prepared and analyzed by EMSA for DNA binding activity as described in panel A (B) or processed for ChIP analysis of Egr-1 binding to FGF-2 promoter in vivo (D).