Innate immunity is the first line of defense against invading pathogens. A family of Toll-like receptors (TLRs) acts as primary sensors that detect a wide variety of microbial components and elicit innate immune responses. All TLR signaling pathways culminate in activation of the transcription factor nuclear factor-kappaB (NF-κB), which controls the expression of an array of inflammatory cytokine genes. NF-κB activation requires the phosphorylation and degradation of inhibitory κB (IκB) proteins, which is triggered by two kinases, IκB kinase α (IKKα) and IKKβ. In addition, several TLRs activate alternative pathways involving the IKK-related kinases TBK1 [TRAF family member-associated NF-κB activator (TANK) binding kinase-1] and IKKi, which elicit antiviral innate immune responses. Here, we review recent progress in our understanding of the role of NF-κB in TLR signaling pathways and discuss potential implications for molecular medicine.