Activation of the transcription factor, NF-κB, during hepatic ischemia/reperfusion injury is associated with proinflammatory mediator expression and is thought to be one of the initial triggers for the inflammatory response after reperfusion. In the current study, we sought to determine whether in vivo adenoviral transfection of a mutant inhibitor of κB-alpha (IκBα), which cannot be serine phosphorylated or degraded (IκBαSR), would inhibit NF-κB and ameliorate the hepatic inflammatory response to ischemia/reperfusion. Male C57BL/6 mice were subjected to sham surgery or partial hepatic ischemia (90 min) and reperfusion (up to 8 h). Mice were infected with 1 × 10 9 PFU of adenovirus containing either β-galactosidase (LacZ) or IκBαSR 3 days prior to induction of ischemia. Serum and tissues were obtained at various times for analysis. In unmanipulated mice, degradation of IκBα, as occurs after serine phosphorylation, was evident in liver by the end of ischemia and during early reperfusion. Mice transfected with IκBαSR displayed the same degree of inflammation and hepatocellular injury as LacZ-transfected mice. There was no difference between LacZ- and IκBαSR-transfected livers in terms of NF-κB activation or proinflammatory cytokine production. The data demonstrate that the pathway of NF-κB activation involving serine phosphorylation of IκBα is not the primary mechanism for induction of liver inflammation after ischemia/reperfusion and suggest that alternative pathways, such as tyrosine phosphorylation of IκBα, may be essential for the postischemic response in liver.