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. 2018 Jul 26;8:11255. doi: 10.1038/s41598-018-29308-7

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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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IH, GN and Aca inhibit PI3K/AKT/mTOR/p70S6K/ULK1 signaling pathway via reducing PI3Kγ-p110 expression. (A)MDA-MB-231 cells were treated with IH, GN, Aca at indicated concentrations for 24 h, cell lysates were prepared and subjected to western blot using antibodies against PI3Kα, PI3Kβ, PI3Kγ, PI3Kδ, p-PI3K, p-AKT, AKT, p-mTOR, mTOR, p-p70S6K, p-ULK and ULK1. (B,C) MDA-MB-231 cells were pretreated with 20 μM AS605240 for 2 h, followed by treatment with or without 20 μM IH, 20 μM GN and 50 μM Aca for 48 h. Cells were stained with Annexin V/PI, and the percentage of apoptotic cells were showed as mean ± S.D. for three independent experiments (**P < 0.01). (D–F) Cells were treated as B and C, cell lysates were prepared and subjected to western blot analysis using antibodies against PARP, cleaved-caspase 3 (C-Caspase 3), Bcl-2, Bcl-xL, p53, PI3Kγ, p-Akt, p-mTOR, p-ULK p70S6K, p62 and LC3.