Abstract
Developmental and immune-mediated disease has been linked to genetic mutation of key signaling components involved in NF-κB activation that leads to impaired activation or regulation of the canonical IKK complex. We identify patients with suspected or known defects of the NF-κB signaling pathway through clinical phenotyping and genetic sequencing. To help understand how mutations cause disease, we quantitate the kinetics and dose-response of NF-κB activation signaling events in their cells. Following activation of the canonical IKK complex, phosphorylation of the inhibitor of NF-κB proteins (IκB) leads to their degradation and the subsequent translocation of NF-κB family members from the cell cytoplasm to the nucleus. Here, we provide a method to obtain patient-derived dermal fibroblasts and quantitatively assess the integrity of the signal transduction pathway from receptor activation to nuclear p65 translocation.
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Wessel, A.W., Hanson, E.P. (2015). A Method for the Quantitative Analysis of Stimulation-Induced Nuclear Translocation of the p65 Subunit of NF-κB from Patient-Derived Dermal Fibroblasts. 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_25
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DOI: https://doi.org/10.1007/978-1-4939-2422-6_25
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2421-9
Online ISBN: 978-1-4939-2422-6
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