Abstract
NF-κB comprises a family of transcription factors that regulate the expression of diverse gene families essential for inflammatory and immune responses as well as cell survival and cell death pathways. Aberrant NF-κB transcriptional activity plays pivotal roles in a large number of human pathologies, including a variety of cancers and chronic inflammatory diseases. Therefore, there has been a large increase in studies aimed at identifying and testing drugs or small molecule inhibitors that would specifically block NF-κB activation in inflammatory diseases and cancer. In this chapter, we describe an in vivo system to test the inhibitory effects of the NEMO-binding domain (NBD) peptide on NF-κB activation specifically in the vascular endothelium and lymphocytes in mice. We demonstrate that pretreatment of mice with the NBD peptide reduces the NF-κB induced gene expression of cell adhesion molecules and DNA-binding activity following systemic LPS stimulation. These methods can be further used to test alternate inhibitors for effects on NF-κB signaling in murine endothelium and immune cells.
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Acknowledgments
Work in the authors’ laboratory was supported by NIH RO1 HL080612 and RO1 HL096642.
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McCorkell, K.A., May, M.J. (2015). NEMO-Binding Domain Peptide Inhibition of Inflammatory Signal-Induced NF-κB Activation In Vivo. 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_30
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DOI: https://doi.org/10.1007/978-1-4939-2422-6_30
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-2422-6
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