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. 2017 Nov 22;7:15992. doi: 10.1038/s41598-017-16149-z

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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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Metformin (Met) does not induce apoptosis, autophagy and senescence in HT29, HCT116 and HCT116 p53−/− cells. (a) Graphic representation of the results obtained from Annexin V assay (upper panel) and western blot analysis of Caspase-3 and PARP activation after treatment with 5 mM Met for 72 h (lower panel). Vinculin was used as the loading control. (b) Immunoblotting determination of LC3B and BECN1 protein after treatment with 5 mM Met for 72 hours; Vinculin was used as the loading control. Full-size blots are shown in Supplementary Fig. S11. (c) β-galactosidase staining of untreated cells (−) and cells treated with Met for 72 hours (+). The images were acquired at 20x magnification. At the bottom, human primary thyrocytes carrying the ER:RAS vector were left untreated or treated with 200 nM 4-hydroxytamoxifen (4OHT) for seven days, and used as positive controls (magnification 40x). The data are representative of at least three independent experiments.