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. 2019 Mar 8;9:3979. doi: 10.1038/s41598-019-40482-0

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© The Author(s) 2019

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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TNMD is involved in glucose metabolism in human differentiated adipocytes. Human adipocytes were transfected with an adenovirus-5 containing a shRNA-TNMD and shRNA-control (scrambled) at day 14 of adipogenesis induction. (a) Glucose transporter 4 (GLUT4) mRNA levels were normalized to those of hypoxanthine-guanine phosphoribosyltransferase-1 (HPRT1), and the data from three independent experiments are presented as the fold-change, which was calculated using the Pfaffl method. (b) GLUT4 protein levels from cell lysates were analyzed by Western blot using a specific antibody against GLUT4, normalized to the internal control (α-tubulin) and expressed as fold-change; the lower section shows a representative crop blot. (c) Glucose uptake levels in shRNA-TNMD-treated adipocytes compared with the shRNA-control or insulin (1 μM, 30 min) as a positive control. (d) Adiponectin (ADIPOQ) mRNA and protein levels expressed as fold-change. (e) Ratio phosphor-AMPKα/total-AMPKα. (f) Ratio phosphor-AKT/total-AKT. (g) Immunofluorescent staining of adipocytes at day 14 with GLUT4 (red) and 4.6-diamidino-2-phenylindole (DAPI; blue; scale bar, 200 μm) in the shRNA-control and shRNA-TNMD-treated adipocytes. All values are expressed as the means ± SEM of three independent experiments. Significant differences were identified using the nonparametric Mann-Whitney U test; *P-value < 0.05.