NEMO-binding domain peptide inhibition of inflammatory signal-induced NF-κB activation in vivo.
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.
Department of Animal Biology, The University of Pennsylvania School of Veterinary Medicine, 3800 Spruce Street (OVH 200E), Philadelphia, PA, 19104, USA.
Kelly A
KA
McCorkell
Michael J
MJ
May
D000818:Animals; D001011:Aorta / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods; D019040:E-Selectin / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D004789:Enzyme Activation / Q000187:drug effects; D007249:Inflammation / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000008:administration & dosage; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D010446:Peptide Fragments / Q000008:administration & dosage / Q000494:pharmacology*; D054730:Protein Interaction Domains and Motifs; D015398:Signal Transduction / Q000187:drug effects*; D013154:Spleen / Q000378:metabolism; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
D019040:E-Selectin; D047908:Intracellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides; C514981:NEMO protein, mouse; D016328:NF-kappa B; D010446:Peptide Fragments; D019010:Vascular Cell Adhesion Molecule-1