Bioluminescence Analysis of Smad-Dependent TGF-β Signaling in Live Mice

  • Jian Luo
  • Tony Wyss-Coray
Part of the Methods in Molecular Biology™ book series (MIMB, volume 574)


TGF-β signaling via the Smad2/3 pathway has key roles in development and tissue homeostasis. Perturbations of the TGF-β signaling are involved in the pathogenesis of many human diseases, including cancer, fibrotic disorders, developmental defects, and neurodegeneration. To study the temporal and spatial patterns of Smad2/3-dependent signaling in living animals, we engineered transgenic mice with a Smad-responsive luciferase reporter (SBE-luc mice). Smad2/3-dependent signaling can be assessed non-invasively in living mice by bioluminescence imaging. To identify the cellular source of the bioluminescence signal, we generated new reporter mice expressing a trifusion protein containing luciferase, red fluorescent protein (RFP), and thymidine kinase under the control of the same SBE promoter (SBE-lucRT mice). SBE-luc and SBE-lucRT mice can be used to study temporal, tissue-specific activation of Smad2/3-dependent signaling in living mice as well as for the identification of endogenous or synthetic modulators of this pathway.

Key words

TGF-β Smad bioluminescence imaging in vivo luciferase immunofluorescence 



We thank B. Debsi, W. Wang, H. Yang, and E. Hashemi for animal husbandry and genotyping. This work was supported by grants from NIH (AG23708, AG20603) and the John Douglas French Alzheimer’s Foundation (T.W-C).


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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Jian Luo
    • 1
  • Tony Wyss-Coray
    • 2
    • 3
  1. 1.Department of Neurology and Neurological SciencesStanford University School of MedicineStanfordUSA
  2. 2.Department of Neurology and Neurological SciencesStanford University School of MedicineStanfordUSA
  3. 3.GRECC, VA Palo Alto Health Care SystemPalo AltoUSA

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