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. 2017 Aug 24;7:9373. doi: 10.1038/s41598-017-09668-2

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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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Combination of NCTD and CLSO induces cell cycle arrest and apoptosis in HepG2 and HepG2/ADM cells. (a) Cell cycle distribution of HepG2 and HepG2/ADM cells was determined 24 h after treatment with NCTD and CLSO alone or in combination(n = 3). (b) Cells were stained with Hoechst33342 (5 μg/ml) and subjected to analysis of apoptosis population(n = 3). (c) PE-Annexin V staining of phosphatidylserine exposed on the cell surface was measured by flow cytometric analysis (n = 3). Data derived from three separate experiments are presented as the means ± S.D. (d) Total cell lysates were prepared for western blot analysis of the apoptosis regulatory proteins (n = 3). ^*P < 0.05; ^**P < 0.01, vs. control, ^aP < 0.05; ^aaP < 0.01 vs. NCTD alone, ^bP < 0.05; ^bbP < 0.01 vs. CLSO alone. One-way ANOVA, post hoc comparisons, Tukey’s test. Columns, means; error bars, SDs.