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. 2019 Jul 23;9:10673. doi: 10.1038/s41598-019-46764-x

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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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Effect of autophagy on the mesenchymal phenotype of ductular cells from AAF/CCL4 livers. (A,B) Immunofluorescence images of isolated ductular cells demonstrating LC3B was associated with the mesenchymal markers vimentin (A) and α-SMA (B). (Right) Quantification of images was performed using Zen-Pro software (Zeiss) and normalized to the number of nuclei. (C) Ductular cells were cultured for 3 days and then treated with or without bafilomycin A1 (Baf, 100 nM) for another 24 hours. Representative Western blot analysis confirmed the increased expression of mesenchymal markers in AAF/CCL4 cells compared to normal cells. Treatment of AAF/CCL4 cells with Baf suppressed the expression of mesenchymal markers. Full-length blots are presented in Supplementary Fig. S7. Quantitative analysis of mesenchymal markers (left panel), and autophagic markers (right panel) was shown below. (D) Inhibition of autophagy by ATG7 small interfering RNA (siATG7) reduced the protein levels of mesenchymal markers. Full-length blots are presented in Supplementary Fig. S8. Quantitative analysis of mesenchymal markers (left panel), and autophagic markers (right panel) was shown below. Statistical analysis was analyzed by Student’s unpaired t-test. *P < 0.05 compared with normal group; ^s#P < 0.05 compared with AAF/CCL4 groups.