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. 2019 Mar 25;9:5039. doi: 10.1038/s41598-019-41527-0

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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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Effect of CFZ on cell cycle arrest and apoptosis in SK-N-BE(2)-M17 cells. (A) SK-N-BE(2)-M17 cells were treated with vehicle or 100–400 nM CFZ for 24 h and then stained with PI and analyzed by flow cytometry. The percentage of gated cells in the subG1, G0/G1, S, and G2/M areas is shown in the upper right area of each plot. (B) Cells were treated with vehicle or 100–400 nM CFZ for 24 h. Vehicle- or CFZ-treated cells were stained with PI and annexin V-FITC and evaluated by FACS analysis. The lower right and upper right areas represent the percentages of early apoptotic cells and late apoptotic cells, respectively. The fraction of necrotic cells is shown in the upper left area. (C) Cells were treated with various concentrations (20–400 nM) of CFZ for 24 h (CFZ-dose) or treated with 200 nM CFZ for the indicated durations (CFZ-time). Cells were then lysed and total cell extracts were resolved by SDS-PAGE. The levels of activated caspases were detected by western blot analysis using antibodies against cleaved forms of caspases (Cas-8, 9, 4, and 3), and β-actin (internal control). Blots are representative of those obtained in more than three independent experiments.