PlGF augments HRE-Luc and FLAP-Luc promoter via activation of PI-3 kinase and HIF-1α. THP-1 cells cotransfected with HRE-Luc and β-galactosidase plasmid were (A) either pretreated with LY294002 or cotransfected with PTEN and (B) cotransfected with indicated plasmids before treatment with PlGF for 24 hours. (C) Deletion analysis of FLAP promoter. THP-1 cells were cotransfected with indicated deletion construct and β-galactosidase plasmid, followed by PlGF treatment for 24 hours. (D) Schematics of FLAP promoter (−371 bp) indicating the location of HIF-1α, NF-κB, and C/EBP binding sites. (E,F) PlGF augments minimal FLAP promoter activity through HREs but not NF-κB in THP-1 (E) and PBM (F). THP-1 cells or PBM were cotransfected with indicated promoter constructs and β-galactosidase plasmid, followed by PlGF treatment for 24 hours. (G) THP-1 cells were treated with either indicated pharmacologic inhibitors or transfected with siRNA or HIF expression plasmids. These cells were then cotransfected with −371-FLAP-Luc and β-galactosidase plasmid, followed by PlGF treatment for 24 hours. Luciferase and β-galactosidase activities were measured as described in “Transient transfection.” The luciferase activity was normalized to that of the promoterless pGL3 basic vector. Data are expressed as mean plus or minus SEM of 3 independent experiments (***P < .001; **P < .01; ns, P > .05).