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. 2017 Jun 9;7:3158. doi: 10.1038/s41598-017-03230-w

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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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TRPC5 induces autophagy via the CaMKKβ/AMPKα/mTOR pathway in response to chemotherapy. (A and B) Treatment of indicated cells with ADM for 48 h increased the basal [Ca²⁺]_i and siTRPC5 blocked the ADM-induced increase in [Ca²⁺]_i. (C) Exposure of indicated cells to ADM for 48 h was followed by analysis of the protein levels of p-CaMKKβ, CaMKKβ, p-AMPKα, AMPKα, p-mTOR, mTOR, p-p70S6K, p70S6K and ACTB by western blot. (D) The effect of TRPC5 silencing on the protein levels of p-CaMKKβ, CaMKKβ, p-AMPKα, AMPKα, p-mTOR, mTOR, p-p70S6K, p70S6K and ACTB in indicated cells exposed to ADM. (E) The effect of 20 µmol/L BAPTA/AM on the protein levels of p-CaMKKβ, CaMKKβ, p-AMPKα, AMPKα, p-mTOR, mTOR, p-p70S6K, p70S6K, LC3 and ACTB in indicated cells exposed to ADM. (F) The effect of CaMKKβ silencing on the protein of CaMKKβ, p-AMPKα, AMPKα, p-mTOR, mTOR, p-p70S6K, p70S6K, LC3 and ACTB in indicated cells treated with ADM. (G) The effect of AMPKα silencing on the protein levels of AMPKα, p-mTOR, mTOR, p-p70S6K, p70S6K, LC3 and ACTB in indicated cells exposed to ADM. (H and I) The effect of BAPTA/AM, siCaMKKβ and siAMPKα on ADM induced LC3 puncta formation in indicated cells. (J and K) The effect of 10 µmol/L STO-609 and 5 µmol/L compound C on ADM induced LC3 puncta formation in indicated cells. (L) The effect of BAPTA/AM, STO-609 and compound C on accumulation of LC3-II in MCF-7/ADM cells. Values are mean ± SEM of 3 to 6 experiments. *p < 0.05, **p < 0.01, compared to siCTL or vehicle.