ABSTRACTLipopolysaccharides (LPS) are potent polyclonal B-lymphocyte activators. Recently, we have shown that LPS inhibits both spontaneous and drug-induced apoptosis in mature B lymphocytes, through cytosolic retention of Bax, a proapoptotic protein of the Bcl-2 family, by preventing its translocation to mitochondria. Research within the last few years has revealed that members of the NF-κB transcription factor regulate cell viability by activating genes involved in mitochondrion-dependent apoptosis. In this report, we examined the effect of sustained LPS stimulation on cytosolic and nuclear proteins of the IκB/NF-κB family to determine which NF-κB pathway, canonical (classical) or noncanonical (alternative), is activated by this agent in mature B cells. Immunoblotting analyses showed that LPS induced a time-dependent degradation of the NF-κB inhibitors IκBβ and IκBε (preferentially to isoform IκBα), via IκB kinase β. In addition, we observed that LPS triggered the processing of NF-κB p105 to p50 and that of NF-κB p100 to p52 in parallel with nuclear translocation of active p50 and p52, as NF-κBp50/RelA and NF-κBp52/RelB heterodimers, respectively. These results suggest that sustained stimulation with LPS can activate NF-κB through both classical and alternative pathways.