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. 2018 Feb 2;8:2277. doi: 10.1038/s41598-018-20768-5

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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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Inhibitory effect of NTS on key adipogenic transcription factors in 3T3-L1 pre-adipocytes. (A) Quantitative real-time PCR was used to quantify the effect of NTS on adipogenic transcription factors PPARγ and C/EBPα. Results are presented as mean ± SD of three independent experiments. Significant difference: ***p < 0.001; **p < 0.01; *p < 0.05 compared with the control group. (B) Western blot analysis showing the effect of NTS on PPARγ and C/EBP α levels. (C) Immunocytochemical assay for PPARγ (adipogenic transcription factor, Red) and DAPI (nucleus stain, Blue). Scale bar = 100 μm. (D) On day 4, adipocyte specific markers (fatty acid synthase, ACC-acetyl-CoA carboxylase, FAT, and SCD-1-stearoyl-CoA desaturase) were analyzed by real-time PCR using mRNA isolated from NTS-treated 3T3-L1 pre-adipocytes. (E) Proteins were extracted and expression levels of FAS, ACC, FABP4, and perilipin were determined by Western blot. Each Western-blotting band represents three experiments performed in triplicate. (F) Immunocytochemical assay for perilipin (lipid droplet marker, green) and DAPI (nucleus stain, blue). Scale bar = 100 μm.