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. 2019 Sep 25;9:13906. doi: 10.1038/s41598-019-50471-y

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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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Effects of PD on autophagic flux in cultured chondrocytes in vitro. (A) TEM images of the autophaghic change in chondrocytes. Single arrow: autophagolysosome and autophagosome with double membrane structure. Original magnification × 30000 (scale bar, 500 nm). (B) Cells transfected with adenovirus harbouring tandem fluorescent mRFP-GFP-LC3 for 24 h were subjected to different treatments. Representative pictures of immunofluorescent chondrocytes expressing mRFP-GFP-LC3. GFP dots are green, and mRFP dots are red. Original magnification × 1600 (scale bar, 20 μm). (C) Semi-quantitative analysis of autophagosomes (AP; yellow dots in merged images) and autolysosomes (AL; red only dots in merged images). (D) Western blot of LC3, Beclin1, and mTOR in different groups were detected respectively and the semi-quantitative analysis of LC3 II/I ratio, Beclin1/GAPDH, and mTOR/GAPDH are shown. (H) Representative pictures of MDC staining. Original magnification × 1600 (scale bar, 20 μm). Values are the means ± SD (n = 5). Bars with different letters are significantly different from each other at p < 0.05 and those with the same letter exhibit no significant difference.