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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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Release of glycerol and lactate to the extracellular medium in 3T3-L1 adipocytes over-expressing Dlk or Notch genes. Relative levels of glycerol released to the extracellular medium in response to isoproterenol from Dlk1 or Dlk2 genes (A), and Notch1, 2, 3 or 4 genes (B) over-expressing adipocytes. (C) Representative microscopy images (400X magnification) of non-transfected (L1C) and transfected 3T3-L1 adipocytes (Empty vectors V1, V2, V3 and V4, and their corresponding over-expressing transfectant) under study. The size of their lipid droplets is showed. Scale bar (80 μm) is shown. Relative levels of lactate in the culture supernatant of differentiated non-transfected 3T3-L1 cells (D), Dlk1 or Dlk2 genes over-expressing adipocytes (E), and each of the Notch genes over-expressing adipocytes (F). The fold activation or inhibition was calculated relative to the seven-day differentiated non-transfected or empty-vector-transfected cells, which was set arbitrarily at 1. Data are shown as the mean ± SD of at least three biological assays performed in triplicate. The statistical significance calculated by Student’s t-tests is indicated (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001).