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. 2019 Dec 27;9:19890. doi: 10.1038/s41598-019-56535-3

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© The Author(s) 2019

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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Induction of autophagy in intestinal epithelial cells. The optimal concentration (A) and time (B) of rapamycin on Caco2 and IEC-6 were respectively detected. Cell lysates were prepared and analyzed by western blotting for the expression of autophagy related proteins LC3, p62 and Beclin1. GAPDH was used as a normalization control. The LC3II/LC3I ratio of Caco2 increased significantly after 24 h incubation with 50 nM rapamycin (C,E), accordingly, 24 h treatment with 200 nM rapamycin could successfully induce autophagy of IEC-6 (D,F). Data was presented as mean ± SD for three independent experiments. **p < 0.01, ***p < 0.001.