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. 2017 Jul 24;7:6248. doi: 10.1038/s41598-017-06576-3

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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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Temporal induction of mTnfaip8 by insulin. (a) mTnfaip8 was overexpressed in mouse hepatoma cells (Hepa 1−6) and phospho-mTOR and LC3 II levels were analyzed by immunoblotting after 48 h. The anti-autophagic activity of mTnfaip8 was confirmed (left panel). Hepa 1-6 cells were treated with insulin (1 µg/ml), and the levels of mTnfaip8 were analyzed by immunoblotting at 0 - 24 h. Insulin induced significant increases in mTnfaip8 levels over 24 h, which was accompanied by a significant decrease in the autophagy marker LC3 II (i.e., inhibition of autophagy) (right panel). (b,c) mTnfaip8 gene KD was performed in either absence or presence of insulin (1 µg/ml). The levels of phospho-PI3K, phospho-Akt and phospho-mTOR were analyzed by immunoblotting after 48 h. The levels of phospho-PI3K, phospho-Akt and phospho-mTOR were decreased significantly and autophagy was induced. Values are means (SD). *p < 0.05, ^##p/**p < 0.01, ^###p/***p < 0.001. The full-length blots are presented in Supplementary Figure 2 (Fig. S2).