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. 2022 Oct 25;20:165. doi: 10.1186/s12964-022-00966-5

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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 — Metformin regulates autophagy through a mechanism involving BECN1 and BNIP3. A Cell lysates of osteoclast precursors were used to examine the expression of BECN1, BNIP3 and LC3 proteins by western bolt. After overexpression of BNIP3 (Lv-BNIP3), osteoclast precursors were treated with metformin (0, 10, 20 μM) for 5 d in the presence of 30 ng/mL M-CSF and 50 ng/mL RANKL. B The quantitative analysis of immunoblots relative to GAPDH protein level in (A). The results were from pairwise comparisons within- and between-group. C Osteoclast precursors were treated as in (A), and the cell lysates were preformed to co-immunoprecipitation with BCL2 antibody, followed by immunoblotting with the indicated antibodies. Among them, IgG group was considered as negative control. D, E Osteoclast precursors were treated as in (A), and the cell lysates were subjected to co-immunoprecipitation with either BCL2 or BECN1 antibody, followed by immunoblotting with the indicated antibodies. Among them, IgG group was considered as negative control. Values displayed are means ± SD of 3 independent experiments (*p < 0.05; **p < 0.01; ***p < 0.001, ns, non-significant)