M. Kerns et al. and the associated impact, represent complex issues (Ma- cLeod et al., 2009). Treatment with SF has been shown to be very effective in alleviating skin blistering in a mouse model for K14 deﬁ- ciency, thus showing promise as a therapeutic agent for a subset of epidermolysis bullosa simplex cases (Kerns et al., 2007). The list of conditions that stand to beneﬁt from treat- ment with natural products enriched in this remarkable molecule (Shapiro et al., 2006; Dinkova-Kostova et al., 2007) is growing steadily. The ﬁndings reported here shed new light into the complexity of the cellular pathways that are impacted by SF and related molecules and as such may assist in efforts to use them appropriately in a clinical setting. ACKNOWLEDGMENTS The authors are grateful to Dr. Paul Talalay (Johns Hopkins School of Med- icine) for advice. These studies were supported by grants to P.A.C. from the March of Dimes Birth Defect Research Foundation and the National Institutes of Health (AR44232, CA123530). REFERENCES Bachelor, M. A., Cooper, S. J., Sikorski, E. T., and Bowden, G. T. (2005). Inhibition of p38 mitogen-activated protein kinase and phosphatidylinositol 3-kinase decreases UVB-induced activator protein-1 and cyclooxygenase-2 in a SKH-1 hairless mouse model. Mol. Cancer Res. 3, 90 –99. Benavides, F. Oberyszyn, T. M., Van Buskirk, A. M., Reeve, V. E., and Kusewitt, D. F. (2009). The hairless mouse in skin research. J. Dermotol. Sci. 53, 10 –18. Chan, K., Lu, R., Chang, J. C., and Kan, Y. W. (1996). NRF2, a member of the NFE2 family of transcription factors, is not essential for murine erythropoie- sis, growth, and development. Proc. Natl. Acad. Sci. USA 93, 13943–13948. Cho, S. D., Li, G., Hu, H., Jiang, C., Kang, K. S., Lee, Y. S., Kim, S. H., and Lu, J. (2005). Involvement of c-Jun N-terminal kinase in G2/M arrest and caspase- mediated apoptosis induced by sulforaphane in DU145 prostate cancer cells. Nutr. Cancer 52, 213–224. Cornblatt, B. S., et al. (2007). Preclinical and clinical evaluation of sulforaphane for chemoprevention in the breast. Carcinogenesis 28, 1485–1490. Coulombe, P. A., Kerns, M. L., and Fuchs, E. (2009). Epidermolysis bullosa simplex: a paradigm for disorders of tissue fragility. J. Clin. Invest. 119, 1784 –1793. Dinkova-Kostova, A. T., Fahey, J. W., Wade, K. L., Jenkins, S. N., Shapiro, T. A., Fuchs, E. J., Kerns, M. L., and Talalay, P. (2007). Induction of the phase 2 response in mouse and human skin by sulforaphane-containing broccoli sprout extracts. Cancer Epidemiol. Biomarkers Prev. 16, 847– 851. Dinkova-Kostova, A. T., Holtzclaw, W. D., Cole, R. N., Itoh, K., Wakabayashi, N., Katoh, Y., Yamamoto, M., and Talalay, P. (2002). Direct evidence that sulfhydryl groups of Keap1 are the sensors regulating induction of phase 2 enzymes that protect against carcinogens and oxidants. Proc. Natl. Acad. Sci. USA 99, 11908 –11913. Endo, H., Sugioka, Y., Nakagi, Y., Saijo, Y., and Yoshida, T. (2008). A novel role of the NRF2 transcription factor in the regulation of arsenite-mediated keratin 16 gene expression in human keratinocytes. Environ. Health Perspect. 116, 873– 879. Fine, J. D., Eady, R. A., Bauer, E. A., Bauer, J. W., Bruckner-Tuderman, L., Heagerty, A., Hintner, H., Hovnanian, A., Jonkman, M. F., Leigh, I., et al., (2008). The classiﬁcation of inherited epidermolysis bullosa (EB): Report of the Third International Consensus Meeting on Diagnosis and Classiﬁcation of EB. J. Am. Acad. Dermatol. 58, 931–950. Hayes, J. D., Kelleher, M. O., and Eggleston, I. M. (2008). The cancer chemo- preventive actions of phytochemicals derived from glucosinolates. Eur. J. Nutr. 47 (Suppl. 2), 73– 88. Heiss, E., Herhaus, C., Klimo, K., Bartsch, H., and Gerha¨user, C. (2001). Nuclear factor kappa B is a molecular target for sulforaphane-mediated anti-inﬂammatory mechanisms. J. Biol. Chem. 276(34), 32008 –32015. Higgins, L. G., Kelleher, M. O., Eggleston, I. M., Itoh, K., Yamamoto, M., and Hayes, J. D. (2009). Transcription factor Nrf2 mediates an adaptive response to sulforaphane that protects ﬁbroblasts in vitro against the cytotoxic effects of electrophiles, peroxides and redox-cycling agents. Toxicol. Appl. Pharmacol. 237, 267–280. 4074 Hu, R., Xu, C., Shen, G., Jain, M. R., Khor, T. O., Gopalkrishnan, A., Lin, W., Reddy, B., Chan, J. Y., and Kong, A. N. (2006). Gene expression proﬁles induced by cancer chemopreventive isothiocyanate sulforaphane in the liver of C57BL/6J mice and C57BL/6J/Nrf2 ( / ) mice. Cancer Lett. 243, 170 –192. Hwang, J. W., Park, J. S., Jo, E. H., Kim, S. J., Yoon, B. S., Kim, S. H., Lee, Y. S., and Kang, K. S. (2005). Chinese cabbage extracts and sulforaphane can protect H2O2-induced inhibition of gap junctional intercellular communica- tion through the inactivation of ERK1/2 and p38 MAP kinases. J. Agric. Food Chem. 53, 8205– 8210. Jaffee, B. D., Manos, E. J., Collins, R. J., Czerniak, P. M., Favata, M. F., Magolda, R. L., Scherle, P. A., and Trzaskos, J. M. (2000). Inhibition of MAP kinase kinase (MEK) results in an anti-inﬂammatory response in vivo. Bio- chem. Biophys. Res. Commun. 268, 647– 651. Jiang, C. K., Flanagan, S., Ohtsuki, M., Shuai, K., Freedberg, I. M., and Blumenberg, M. (1994). Disease-activated transcription factor: allergic reac- tions in human skin cause nuclear translocation of STAT-91 and induce synthesis of keratin K17. Mol. Cell. Biol. 14, 4759 – 4769. Jo, E. H., Kim, S. H., Ahn, N. S., Park, J. S., Hwang, J. W., Lee, Y. S., and Kang, K. S. (2007). Efﬁcacy of sulforaphane is mediated by p38 MAP kinase and caspase-7 activations in ER-positive and COX-2-expressed human breast can- cer cells. Eur. J. Cancer Prev. 16, 505–510. Kang, K. A., Lee, K. H., Park, J. W., Lee, N. H., Na, H. K., Surh, Y. J., and You, H. J., Chung, M. H., Hyun, J. W. (2007). Triphlorethol-A induces heme oxygenase-1 via activation of ERK and NF-E2 related factor 2 transcription factor. FEBS Lett. 581, 2000 –2008. Kerns, M. L., DePianto, D., Dinkova-Kostova, A. T., Talalay, P., and Cou- lombe, P. A. (2007). Reprogramming of keratin biosynthesis by sulforaphane restores skin integrity in epidermolysis bullosa simplex. Proc. Natl. Acad. Sci. USA 104, 14460 –14465. Kim, S., and Coulombe, P. A. (2007). Intermediate ﬁlament scaffolds fulﬁll mechanical, organizational, and signaling functions in the cytoplasm. Genes Dev. 21, 1581–1597. Kim, S., Wong, P., and Coulombe, P. A. (2006). A keratin cytoskeletal protein regulates protein synthesis and epithelial cell growth. Nature 441, 362–365. Kobayashi, M., and Yamamoto, M. (2006). Nrf2-Keap1 regulation of cellular defense mechanisms against electrophiles and reactive oxygen species. Adv. Enzyme Regul. 46, 113–140. Kraft, A. D., Johnson, D. A., and Johnson, J. A. (2004). Nuclear factor E2- related factor 2-dependent antioxidant response element activation by tert- butylhydroquinone and sulforaphane occurring preferentially in astrocytes conditions neurons against oxidative insult. J. Neurosci. 24, 1101–1112. Ku, N. O., and Omary, M. B. (2006). A disease- and phosphorylation-related nonmechanical function for keratin 8. J. Cell Biol. 174, 115–125. Lee, J. C., Kundu, J. K., Hwang, D. M., Na, H. K., and Surh, Y. J. (2007). Humulone inhibits phorbol ester-induced COX-2 expression in mouse skin by blocking activation of NF-kappaB and AP-1, IkappaB kinase and c-Jun-N- terminal kinase as respective potential upstream targets. Carcinogenesis 28, 1491–1498. Leigh, I. M., Navsaria, H., Purkis, P. E., McKay, I. A., Bowden, P. E., and Riddle, P. N. (1995). Keratins (K16 and K17) as markers of keratinocyte hyperproliferation in psoriasis in vivo and in vitro. Br. J. Dermatol. 133, 501–511. Limon-Pacheco, J. H., Hernandez, N. A., Fanjul-Moles, M. L., and Gonsebatt, M. E. (2007). Glutathione depletion activates mitogen-activated protein kinase (MAPK) pathways that display organ-speciﬁc responses and brain protection in mice. Free Radic. Biol. Med. 43, 1335–1347. Ma, S., Rao, L., Freedberg, I. M., and Blumenberg, M. (1997). Transcriptional control of K5, K6, K14, and K17 keratin genes by AP-1 and NF-kappaB family members. Gene Expr. 6, 361–370. MacLeod, A. K., McMahon, M., Plummer, S. M., Higgins, L. G., Penning, T. M., Igarashi, K., and Hayes, J. D. (2009). Characterization of the cancer chemopreventive NRF2-dependent gene battery in human keratinocytes: demonstration that the KEAP1-NRF2 pathway, and not the BACH1-NRF2 pathway, controls cytoprotection against electrophiles as well as redox-cy- cling compounds. Carcinogenesis 30, 1571–1580. Mathew, J., Galarneau, L., Loranger, A., Gilbert, S., and Marceau, N. (2008). Keratin-protein kinase C interaction in reactive oxygen species-induced he- patic cell death through mitochondrial signaling. Free Radic. Biol. Med. 45, 413– 424. McGowan, K. M., and Coulombe, P. A. (1998a). Onset of keratin 17 expression coincides with the deﬁnition of major epithelial lineages during skin devel- opment. J. Cell Biol. 143, 469 – 486. McGowan, K. M., and Coulombe, P. A. (1998b). The wound repair associated keratins 6, 16, and 17: Insights into the role of intermediate ﬁlaments in Molecular Biology of the Cell