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. 2018 Nov 16;8:16923. doi: 10.1038/s41598-018-35252-3

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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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NOTCH activation and signaling in 3T3-L1 cells stably over-expressing each one of the four NOTCH receptors. (A) qRT-PCR analysis of the relative Hes1 and Hey1 mRNA expression levels in the stable Notch1 gene transfectant (L1-N1), the stable Notch2 gene transfectant (L1-N2), the stable Notch3 gene transfectant (L1-N3), and the stable Notch4 gene transfectant (L1-N4). (B) NOTCH transcriptional activity, as measured by gene reporter luciferase assays, in these four Notch genes stable transfectants. (C) qRT-PCR analysis of the relative individual Notch mRNA expression levels in stable Notch1 gene transfectant (L1-N1), the stable Notch2 gene transfectant (L1-N2), the stable Notch3 gene transfectant (L1-N3), and the stable Notch4 gene transfectant (L1-N4). The relative luciferase activities were always normalized with renilla values and referred to those of control cells. Data in all qRT-PCR assays were normalized to P0 mRNA expression levels. The fold activation or inhibition in all assays is measured relative to the empty vector control, set arbitrarily at 1. Data are shown as the mean ± SD of at least three biological assays performed in triplicate. The statistical significance of Student’s t-tests results is indicated (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001).