Abstract
Luciferase reporter assays are sensitive and accurate tests that enable the analysis of regulatory sequences, the magnitude of transcriptional activity by transcription factors, and the discovery of gene regulatory elements and small-molecule modulators with high levels of precision. This is made possible through detection of bioluminescence produced by luciferase-coding reporters in a wide range of cellular environments. These assays are routinely used to analyze the activity of transcription factors, including the lipid-activated liver X receptor (LXR), in response to different stimuli as well as for the identification of their ligands. In this chapter we describe in detail the assays performed to investigate LXR activity in a macrophage-like cell line (RAW 267.4). These can be easily adapted to other nuclear receptors and transcription factors.
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De Wet JR, Wood KV, DeLuca M, Helinski DR, Subramani S (1987) Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol 7:725–737
Hong C, Tontonoz P (2014) Liver X receptors in lipid metabolism: opportunities for drug discovery. Nat Rev Drug Discov 6:433–444
Nwachukwu JC, Li W, Pineda-Torra I, Huang HY, Ruoff R, Shapiro E, Taneja SS, Logan SK, Garabedian MJ (2007) Transcriptional regulation of the androgen receptor cofactor androgen receptor trapped clone-27. Mol Endocrinol 21:2864–2876
Pourcet B, Pineda-Torra I, Derudas B, Staels B, Glineur C (2010) SUMOylation of human peroxisome proliferator-activated receptor alpha inhibits its trans-activity through the recruitment of the nuclear corepressor NCoR. J Biol Chem 285:5983–5992
Pourcet B, Feig JE, Vengrenyuk Y, Hobbs AJ, Kepka-Lenhart D, Garabedian MJ, Morris SM Jr, Fisher EA, Pineda-Torra I (2011) LXRα regulates macrophage arginase 1 through PU.1 and interferon regulatory factor 8. Circ Res 109:492–501
Costet P, Luo Y, Wang N, Tall AR (2000) Sterol-dependent transactivation of the ABC1 promoter by the liver X receptor/retinoid X receptor. J Biol Chem 275:28240–28245
Collins JL, Fivush AM, Watson MA, Galardi CM, Lewis MC, Moore LB, Parks DJ, Wilson JG, Tippin TK, Binz JG, Plunket KD, Morgan DG, Beaudet EJ, Whitney KD, Kliewer SA, Willson TM (2002) Identification of a nonsteroidal liver X receptor agonist through parallel array synthesis of tertiary amines. J Med Chem 45:1963–1966
Acknowledgement
The authors would like to thank the support of the Medical Research Council (New Investigator Grant G0801278), International Reintegration Program (Grant PIRG02-GA-2007-224867) and British Heart Foundation (PG/13/10/30000) to IPT.
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Gage, M.C., Pourcet, B., Pineda-Torra, I. (2016). Luciferase Reporter Assays to Assess Liver X Receptor Transcriptional Activity. In: Waugh, M. (eds) Lipid Signaling Protocols. Methods in Molecular Biology, vol 1376. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3170-5_7
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DOI: https://doi.org/10.1007/978-1-4939-3170-5_7
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3169-9
Online ISBN: 978-1-4939-3170-5
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