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. 2022 Aug 12;20:117. doi: 10.1186/s12958-022-00991-6

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6 — Sirt1 knockdown blocked the upregulation of melatonin on autophagy and mitophagy. A Expression levels of SIRT1 and AMPK in TM3 cells were detected by Western blot and their gray values were analyzed; B The expression levels of autophagy-related proteins Beclin-1, Atg7, Atg5 and LC3 in TM3 cells were detected by Western blot, and the gray values of protein bands were statistically analyzed; C-F. Mitophagy in TM3 cells was marked by COXIV and LC3 antibody co-staining. The fluorescence intensity and the percentage of co-stained cells (counting randomly 150 cells in six visual fields) were statistically analyzed. G-J Immunofluorescent co-staining of BNIP3L and LC3 antibodies in TM3 cells. The fluorescence intensity (randomly counting 100 cells in 10 visual fields) and the percentage of co-stained particles in the cells (totally 30 cells) were statistically analyzed. K The protein level of BNIP3L in TM3 cells was detected by Western blot, and the gray values of the protein bands were analyzed statistically. NG, normal glucose control group; HG, high glucose treatment group; HG + MT, high glucose and melatonin treatment group; siSirt1, HG + MT + siSirt1 group. A-B, Data are expressed as fold change relative to NG. *, P < 0.05; #, P < 0.01. Bar: C, 100 μm; G, 10 μm