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. 2017 Apr 6;7:672. doi: 10.1038/s41598-017-00796-3

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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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NaF-induced apoptosis in TCMK-1 cells. Cells were incubated with the indicated doses of NaF for 12 h. (a) Representative images of flow cytometry by Annexin V-FITC/PI dual staining. (b) Apoptotic cells are represented as the percentage of Annexin-V single positive plus Annexin-V/PI double-positive cells. (c) Caspase-3 activity was measured using a caspase-3 fluorescent assay kit. Cells were harvested and lysed to detect the cytoplasmic (d) and mitochondrial (e) levels of Bax and Bcl-2. (f) The levels of AIF and CytC in the cytosolic fraction were detected in TCMK-1 cells. All results are representative of three independent experiments and values are presented as means ± s.d. ^*p < 0.05, ^**p < 0.01 versus the control group. Full-length blots/gels are presented in Supplementary Figures S9–11.