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. 2017 Jul 11;7:5040. doi: 10.1038/s41598-017-05052-2

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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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Metformin induces ER stress and calcium release from the ER. (A) List of genes related to the ER stress pathway and significantly upregulated in LNCaP cells treated with 5 mM metformin for 14 h. (B) Immunoblotting of LNCaP cells treated with 5 mM metformin for the indicated times by using antibodies directed against the indicated proteins implicated in ER stress response. The immunoblot is representative of three independent experiments. The blots represent (C) Relative fluorescence intensity of the Fluo-4AM probe, measured in LNCaP cells. 2-ABP (40 µM) was added 15 min before the addition of 5 mM metformin at time 0. The cellular area was monitored during the experiment (blue line). At least 30 single cells were monitored for each condition. The graph represents the mean fluorescence intensity +/− sem. (D) Quantification of IP3R mRNA in LNCaP cells treated with metformin for 24 h. p < 0.05 (*); p < 0.01 (**).