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. 2018 Jul 19;8:10923. doi: 10.1038/s41598-018-29199-8

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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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Compound E5 induced apoptosis in HCC cells. (A) Apoptotic nuclei manifested condensed or fragmented DNA that were brightly stained by Hoechst 33258 (24 h). Magnification, × 200. (B) Flow cytometry analysis after AnnexinV-FITC/PI double staining. Representative histograms for each treatment are shown. (C) Western blot analysis of the caspase cascade and apoptosis related proteins after treatment with E5 for 24 h in Bel7404 and HepG2 cells. Original gel images are presented in Supplementary Fig. S1. (D) Pan-caspase inhibitor Z-VAD-FMK rescued HCC cells from E5-reduced cell viability. HCC cells were exposed to Z-VAD-FMK (10 μM) with or without E5 (20 μM) for 24 h, cell viability was measured by MTT assay. The data were presented as mean ± SD. *P < 0.05; **P < 0.01 compared to DMSO control; one-way ANOVA, post-hoc intergroup comparisons, Tukey’s test.