15(S)-HETE–induced FGF-2 promoter-luciferase expression requires Egr-1-binding site. (A) Sequence of a 0.5-kb cloned human FGF-2 promoter region showing the Egr-1 binding sites. Sequence in bold represents Egr-1-binding site; +1 indicates transcription start site. (B) HDMVECs were transfected with either empty vector or FGF-2 promoter-luciferase constructs, quiesced, and treated with and without 0.1μM 15(S)-HETE for 30 minutes, and cell extracts were prepared and analyzed for luciferase activity. The bar graph in panel B represents the quantitative analysis of 3 independent experiments. The values are presented as the means ± SD. *P < .01 vs hFGF2p-WT-Luc vehicle control; **P < .01 vs hFGF2p-WT-Luc + 15(S)-HETE. hFGF2p-WT-Luc indicates human FGF-2 promoter-luciferase construct; hFGF2p-M1-Luc, human FGF-2 promoter-luciferase construct with mutation in the first Egr-1 binding site from the transcriptional start site; hFGF2p-M2-Luc, human FGF-2 promoter-luciferase construct with mutation in the second Egr-1 binding site from the transcriptional start site; hFGF2p-DM-Luc, human FGF-2 promoter-luciferase construct with mutations in both the Egr-1 binding sites.