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. 2018 Jul 27;8:11377. doi: 10.1038/s41598-018-29063-9

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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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Effect of annatto-extracted tocotrienols on immunohistochemical and TUNEL analysis of pancreatic tissue. Pancreatic tissue sections collected from mice fed LFD (A and F), HFD (B and G), and HFD supplemented with T400 (C and H), T1600 (D and I) or Met (E and J) were immunostained for insulin (A–E) or glucagon (F–J) and counterstained with hematoxylin. Pancreatic tissue sections collected from mice fed LDF (K) or HFD (L) were analyzed for apoptosis by TUNEL assay (green color, K and L). Sections were counterstained with DAPI (blue color, K and L). Total numbers of apoptotic cells from LFD (grey bar) or HFD (black bar) groups (M) were shown. Data shown are means ± SEM (n = 4). Significant differences in means were determined by the unpaired Student t-test method at a P value of ≤0.05.