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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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The proteins DLK1 and DLK2 inhibit the activation of each one of the four NOTCH receptors. NOTCH transcriptional activity, as measured by gene reporter luciferase assays, was determined in Balb/c14 cells transiently co-transfected with a plasmid driving the expression of each NOTCH receptor: N1 (A), N2 (B), N3 (C) and N4 (D); and/or a plasmid driving the expression of DLK1 or DLK2 extracellular regions (A–D). (E) Analysis of NOTCH transcriptional activity in Balb/c14 cells treated with DMSO or with the inhibitor DAPT (10 micromolar). The fold-activation or inhibition in these luciferase assays is measured relative to the control (C), 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).