Abstract
The transcription factor NF-κB regulates expression of a diverse set of genes to modulate multiple biological and pathological processes. Among these, NF-κB activation in response to genotoxic agents has received considerable attention due to its role in regulating cancer cell resistance to chemo- and radiation therapy. Furthermore, induction of this pathway by endogenous damage is further implicated in normal developmental processes, such as B cell development, and premature aging, among others. This pathway also serves as a signaling model in which nuclear initiated signals (DNA damage) are communicated to a cytoplasmic target (IκB kinase and NF-κB). Several of the critical molecular events of this nuclear to cytoplasmic NF-κB signaling cascade were discovered, in part, by genetic complementation analyses of the NEMO-deficient 1.3E2 mouse pre-B cell line. This chapter describes methods used to generate and analyze such reconstitution cell systems and certain caveats that are critical for proper interpretation of NEMO mutant defects.
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Acknowledgements
The authors would like to thank the many Miyamoto lab members, past and present, for their help in formulating this chapter. We would like to particularly thank Shelly Wuerzberger-Davis who has trained generations of lab members and who made significant contributions to this protocol. This work was funded by F30CA171840 (S.S.J.), NIH R01CA077474 (S.M.), NIH R01GM083681 (S.M.).
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Jackson, S.S., Miyamoto, S. (2015). Dissecting NF-κB Signaling Induced by Genotoxic Agents via Genetic Complementation of NEMO-Deficient 1.3E2 Cells. In: May, M. (eds) NF-kappa B. Methods in Molecular Biology, vol 1280. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2422-6_11
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DOI: https://doi.org/10.1007/978-1-4939-2422-6_11
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