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. 2018 Mar 2;9(3):353. doi: 10.1038/s41419-018-0383-6

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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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Fig. 3 — MCF-7 cells were treated with IC₅₀ dose of C10 or vehicle for indicated time. a, d Representative immunofluorescence images of cells stained for LC3B (green) in (a) or ATG16L (red) in (d); nuclei stained with DAPI (blue) (one of 2 or 3 independent experiments for ATG16L and LC3B, respectively). Scale bars: 20 µm. b Quantification of LC3B foci per cell performed using immunofluorescence microscopy. Foci were counted in at least 350 cells in each of 3 independent experiments. The data represent median and IQR. ***P < 0.001 vs. control. c Quantitative analysis of cell area occupied by autophagic vesicles in control cells and cells treated with IC₅₀ dose of C10 based on electron microscopy images. The surface of vesicles in at least 30 cells per sample was counted. The data represent median and IQR. ***P < 0.001 vs. control. e, f Representative western blots showing protein level of (e) p62/SQSTM1, ATG5, LC3B and (f) phospho-AKT (p-AKT), AKT, phospho-p70S6K (p-p70S6K)