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. 2018 Mar 29;8:5376. doi: 10.1038/s41598-018-23698-4

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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The T-REx assay system. Without Dox, expression of Notch1(1704–2531) is suppressed by TetR. The addition of Dox inactivates TetR to initiate expression of Notch1(1704–2531), which is then cleaved by endogenous γ-secretase to form NICD. NICD then translocates to the nucleus and forms a heterotrimer with RBP-J and MAML that binds to a 12× RBP-J binding site to drive luciferase expression. Luciferase activity was detected as Notch signaling pathway activity. Dox: doxycycline, TetR: tetracycline repressor, Luc.: luciferase.