Abstract
Identification and verification of novel transcription factor interactions is an inherent step in the discovery of molecular mechanisms driving gene transcription and regulation. Co-immunoprecipitation and GST-pull down are often key techniques in the verification process. Despite wide applicability, their use may sometimes be restricted. We provide a detailed protocol for an intracellular immunofluorescence technique that may be used as an alternative or complimentary study for transcription factor interaction verification.
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References
Miller J, Stagljar I (2004) Using the yeast two-hyrbid system to identify interacting proteins. Methods Mol Biol 261:247–262
Vikis HG, Guan KL (2004) Glutathione-S-transferase-fusion based assays for studying protein-protein interactions. Methods Mol Biol 261:175–186
Cekan SZ (2002) Genes and transcription factors, including nuclear receptors: methods of studying their interactions. J Lab Clin Med 140:215–227
Mackay JP, Sunde M, Lowry JA, Crossley M, Matthews JM (2007) Protein interactions: is seeing believing? Trends Biochem Sci 32: 530–531
Boogerd CJ, Wong LY, van den Boogaard M, Bakker ML, Tessadori F, Bakkers J, ’t Hoen PA, Moorman AF, Christoffels VM, Barnett P (2011) Sox4 mediates Tbx3 transcriptional regulation of the gap junction protein Cx43. Cell Mol Life Sci 68:3949–3961
Farin HF, Bussen M, Schmidt MK, Singh MK, Schuster-Gossler K, Kispert A (2007) Transcriptional repression by the T-box proteins Tbx18 and Tbx15 depends on Groucho corepressors. J Biol Chem 282:25748–25759
Acknowledgments
This work was supported by European Community’s Sixth Framework Programme contract HeartRepair LSHM-CT-2005-018630.
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Boogerd, C.J.J., Christoffels, V.M., Barnett, P. (2013). Nuclear Recruitment Assay as a Tool to Validate Transcription Factor Interactions in Mammalian Cells. In: Bina, M. (eds) Gene Regulation. Methods in Molecular Biology, vol 977. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-284-1_19
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DOI: https://doi.org/10.1007/978-1-62703-284-1_19
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-283-4
Online ISBN: 978-1-62703-284-1
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