Activation of the transcription factor NF-κB is critical for the tumor necrosis factor-α (TNF-α)-induced inflammatory response. Here we report the complete gene expression profile from activated microvascular endothelial cells emphasizing the direct contribution of the NF-κB pathway. Human microvascular endothelial cell line-1 (HMEC-1) cells were modified to express dominant interfering mutants of the IKK/NF-κB signaling module and expression profiles were determined. Our results provide compelling evidence for the virtually absolute dependence of TNF-α-regulated genes on NF-κB. A constitutively active IKK2 was sufficient for maximal induction of most target genes, whereas a transdominant IκBα suppressed gene expression. Several genes with a critical role in atherogenesis were identified. The endothelial lipase (EL) gene, a key enzyme involved in lipoprotein metabolism, was investigated more in detail. Binding sites interacting with NF-κB in vitro and in vivo were identified and co-transfection experiments demonstrated the direct regulation of the EL promoter by NF-κB. We conclude that targeting the IKK/NF-κB pathway or specific genes downstream may be effective for the control or prevention of chronic inflammatory diseases such as atherosclerosis.