Abstract
The antioxidant response element (ARE) signaling pathway plays an important role in the amelioration of cellular oxidative stress. Thus, assays that detect this pathway can be useful for identifying chemicals that induce or inhibit oxidative stress signaling. The focus of this chapter is to describe a cell-based ARE assay in a quantitative high-throughput screening (qHTS) format to test a large collection of compounds that induce nuclear factor erythroid 2-related factor (Nrf2)/ARE signaling. The assay is described through cell handling, assay preparation, and instrument usage.
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References
Hu R, Saw CL, Yu R, Kong AN (2010) Regulation of NF-E2-related factor 2 signaling for cancer chemoprevention: antioxidant coupled with antiinflammatory. Antioxid Redox Signal 13(11):1679–1698. doi:10.1089/ars.2010.3276
Saw CL, Kong AN (2011) Nuclear factor-erythroid 2-related factor 2 as a chemopreventive target in colorectal cancer. Expert Opin Ther Targets 15(3):281–295. doi:10.1517/14728222.2011.553602
Tufekci KU, Civi Bayin E, Genc S, Genc K (2011) The Nrf2/ARE pathway: a promising target to counteract mitochondrial dysfunction in Parkinson’s disease. Parkinsons Dis 2011:314082. doi:10.4061/2011/314082
Al-Dalaen SM (2014) Review article: oxidative stress versus antioxidants. Am J Biosci Bioeng 2(5):60. doi:10.11648/j.bio.20140205.11
Kensler TW, Wakabayashi N, Biswal S (2007) Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway. Annu Rev Pharmacol Toxicol 47:89–116. doi:10.1146/annurev.pharmtox.46.120604.141046
Nguyen T, Nioi P, Pickett CB (2009) The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem 284(20):13291–13295. doi:10.1074/jbc.R900010200
Shukla SJ, Huang R, Simmons SO, Tice RR, Witt KL, Vanleer D, Ramabhadran R, Austin CP, Xia M (2012) Profiling environmental chemicals for activity in the antioxidant response element signaling pathway using a high throughput screening approach. Environ Health Perspect 120(8):1150–1156. doi:10.1289/ehp.1104709
Wu RP, Hayashi T, Cottam HB, Jin G, Yao S, Wu CC, Rosenbach MD, Corr M, Schwab RB, Carson DA (2010) Nrf2 responses and the therapeutic selectivity of electrophilic compounds in chronic lymphocytic leukemia. Proc Natl Acad Sci U S A 107(16):7479–7484. doi:10.1073/pnas.1002890107
Zlokarnik G, Negulescu PA, Knapp TE, Mere L, Burres N, Feng L, Whitney M, Roemer K, Tsien RY (1998) Quantitation of transcription and clonal selection of single living cells with beta-lactamase as reporter. Science 279(5347):84–88
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Zhao, J., Shukla, S.J., Xia, M. (2016). Cell-Based Assay for Identifying the Modulators of Antioxidant Response Element Signaling Pathway. In: Zhu, H., Xia, M. (eds) High-Throughput Screening Assays in Toxicology. Methods in Molecular Biology, vol 1473. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6346-1_6
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DOI: https://doi.org/10.1007/978-1-4939-6346-1_6
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