Understanding the molecular pathway(s) controlling the expression of stromal cellular antioxidants and phase 2 enzymes is of importance for developing strategies to protect against bone marrow toxicity induced by oxidants and electrophiles. Accordingly, this study was undertaken to determine the role of the nuclear factor E2-related factor 2 (Nrf2) in regulation of both constitutive and chemoprotectant-inducible expression of antioxidants and phase 2 enzymes in mouse bone marrow stromal cells. The constitutive expression of a series of antioxidants and phase 2 enzymes was significantly lower in stromal cells derived from Nrf2 knockout (Nrf2(-/-)) mice than those from wild-type littermates (Nrf2(+/+)). Incubation of Nrf2(+/+) stromal cells with 3H-1,2-dithiole-3-thione (D3T) led to a significant induction of various antioxidants and phase 2 enzymes. The inducibility of the above cellular defenses by D3T was abolished in Nrf2(-/-) cells. As compared to wild-type cells, Nrf2(-/-) cells were much more susceptible to cytotoxicity induced by reactive oxygen or nitrogen species, 4-hydroxy-2-nonenal, 1,4-hydroquinone, or 1,4-benzoquinone. Upregulation of the antioxidants and phase 2 enzymes by D3T in Nrf2(+/+) stromal cells resulted in increased resistance to the above oxidant- and electrophile-induced cytotoxicity, whereas D3T treatment of Nrf2(-/-) cells only provided a marginal cytoprotection. Taken together, this study demonstrates that Nrf2 is crucial in controlling the expression of bone marrow stromal antioxidants and phase 2 enzymes as well as the susceptibility of these cells to oxidative and electrophilic stress.