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. 2018 Sep 21;8:14161. doi: 10.1038/s41598-018-32256-x

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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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Autophagy inhibition synergizes with Vps34 inhibition or macropinocytosis inhibition to selectively inhibit proliferation of Tsc2-deficient cells. (A) Immunoblots confirmed Tsc2 downregulation in Atg5^+/+ and Atg5^−/− MEFs. The blot was cropped to highlight the relevant bands. The full-length blot is presented in Supplementary Figure 5A. (B) Proliferation of Atg5^−/− shTsc2 MEFs grown in 1% FBS DMEM was selectively inhibited by Vps34 inhibitor SAR405 (2 uM, 48 hours). (C) Vps34 downregulation sensitized Tsc2^−/− MEFs to CQ treatment (5 uM, 72 hours). (D) Proliferation of Atg5^−/− shTsc2 MEFs grown in 1% FBS DMEM was selectively inhibited by the macropinocytosis inhibitor EIPA (6 uM, 48 hours). Data represented as mean +/− standard deviation of six biological replicates. Statistical significance was assessed using two-way and one-way ANOVAs with Bonferroni correction with *p < 0.05, ***p < 0.001, ****p < 0.0001. ^ƚp < 0.05, ^ƚƚƚƚp < 0.0001 when comparing treated to vehicle treatment in same cell type.