NF-κB Transcription Factors: Methods and Protocols

Part I: General Methods for Analyzing NF-κB Activation

1. Measuring NF-kB Phosphorylation and Acetylation

Nikita Tushar Modi and Lin-Feng Chen

2. Biochemical Methods to Analyze the Subcellular Localization of NF- κB Proteins Using Cell Fractionation

Davide Vecchiotti, Daniela Verzella, Daria Capece, Mauro Di Vito Nolfi, Barbara Di Francesco, Jessica Cornice, Guido Franzoso, Edoardo Alesse, and Francesca Zazzeroni

3. Immunohistochemical Analysis of Expression, Phosphorylation, and Nuclear Translocation of NF-κB Proteins in Human Tissues

Davide Vecchiotti, Daniela Verzella, Daria Capece, Jessica Cornice, Mauro Di Vito Nolfi, Barbara Di Francesco, Guido Franzoso, Edoardo Alesse, and Francesca Zazzeroni

4. High-Throughput Analysis of the Cell- and DNA Site-Specific Binding of Native NF-κB Dimers Using Nuclear Extract Protein-Binding Microarrays (NextPBMs)

Heather Hook, Rose Zhao, David Bray, Jessica L. Keenan, and Trevor Siggers

5. Molecular and Biochemical Approaches to Study the Evolution of NF-κB Signaling in Basal Metazoans

Pablo J. Aguirre Carri鏮, Leah M. Williams, and Thomas D. Gilmore

Part II: Methods for Studying the Activation of NF-κB Downstream of Distinct Signaling Pathways

6. Methods for Modulating the Pathway of NF-kB Using Short Hairpin RNA (ShRNA)

Marta Moretti, Barbara Di Francesco, Mauro Di Vito Nolfi, Annapaola Angrisani, and Enrico De Smaele

7. Immunoblot Analysis of the Regulation of TNF Receptor Family-Induced NF-kB Signaling by c-IAP Proteins

Eugene Varfolomeev, Tatiana Goncharov, and Domagoj Vucic

8. Methods to Study CARD11-BCL10-MALT1 Dependent Canonical NF-kB Activation in Jurkat T-Cells

Andreas Gewies, Carina Gra , and Daniel Krappmann

9. Analysis of Calcium Control of Canonical NF-κB Signaling in B Lymphocytes

Corbett T. Berry, Michael J. May, and Bruce D. Freedman

10. A Kinase Assay for Measuring the Activity of the NIK-IKK1 Complex Induced via the Noncanonical NF-κB Pathway

Tapas Mukherjee, Yashika Ratra, Balaji Banoth, Alvina Deka, Smarajit Polley, and Soumen Basak

11. Analyze the SUMOylation of IKKγ/NEMO During Genotoxic Stress

Zhao-Hui Wu and Shigeki Miyamoto

Part III: Methods for Analyzing NF-κB Activation in Physiology and Disease

12. Analysis of the Contribution of NF-kB in the Regulation of Chemotherapy-Induced Cell Senescence by Establishing a Tetracycline-Regulated Cell System

Francesco Pacifico, Elvira Crescenzi, and Antonio Leonardi

13. Methods to Detect NF-κB Activity in Tumor-Associated Macrophage (TAM)