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. 2017 Aug 25;7:9536. doi: 10.1038/s41598-017-10209-0

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© The Author(s) 2017

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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ROS induces cholesterol accumulation. (a) INS-1 cells were treated with AAPH in concentrations from 0 to 5 mg/ml for 4 h in serum-free media. Free cholesterol was measured using filipin. *p < 0.05 against control (no AAPH). Scale bars, 10 μm. (b) INS-1 cells were treated with 2.5 mg/ml AAPH for the different periods of time. Cellular staining of carboxy-H₂DFFDA for ROS content and filipin for free cholesterol content was quantified. *p < 0.05 against control (no treatment). (c) Filipin staining of INS-1 cells incubated for 1 h with 5 mg/ml AAPH in serum free media. Scale bars, 50 μm. Filipin intensity per cell was quantified. *p < 0.05. (d) INS-1 cells were treated with LG, HGP, HGP + NAC (0.5 mM), or HGP + Mev for 48 h. Cells were imaged with filipin to measure free cholesterol. Scale bars, 20 μm. *p < 0.05 against HGP.