Abstract
The E2F transcription factors are key targets for the retinoblastoma (RB) tumor suppressor function. The active or inactive status of RB determines the degree by which E2F-dependent gene expression will occur in a given condition. Changes in transcriptional activity in response to extracellular or intracellular stimuli are frequently measured using genetic reporter assays. In particular, dual luciferase reporter assays are most recommended for this purpose because of their improved experimental accuracy. Here we illustrate the usefulness of the dual luciferase reporter assay to detect E2F-mediated transcriptional activity upon overexpression of E2F1 in cultured cells as readout for RB status and function.
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Acknowledgments
This work was supported by grants from the Spanish Ministry (SAF2015-67562-R, cofunded by the European Regional Development fund) and the Basque Government (IT634-13 and KK-2015/89) to A.M.Z.
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Iglesias-Ara, A., Osinalde, N., Zubiaga, A.M. (2018). Detection of E2F-Induced Transcriptional Activity Using a Dual Luciferase Reporter Assay. In: Santiago-Cardona, P. (eds) The Retinoblastoma Protein. Methods in Molecular Biology, vol 1726. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7565-5_14
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DOI: https://doi.org/10.1007/978-1-4939-7565-5_14
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