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. 2020 Jul 3;11:269. doi: 10.1186/s13287-020-01785-6

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© The Author(s) 2020

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. 3 — HDAC9 regulated the autophagy in BMMSCs by directly binding to the promoters of autophagy-related genes. To evaluate the role of HDAC9 in regulating autophagy, young and aged BMMSCs cultured in vitro and aged BMMSCs were transfected with Nc siRNA or HDAC9 siRNA. a LC3 was measured by immunofluorescence staining in young and aged BMMSCs, and those cells treated with CQ. Scale bars = 50 μm. b Autophagy-related proteins were detected in young and aged BMMSCs, and those cells treated with CQ by western blotting. c LC3 was measured by immunofluorescence staining in aged BMMSCs transfected with Nc siRNA or HDAC9 siRNA, and those cells treated with CQ. Scale bar = 50 μm. d Autophagy-related proteins were detected in aged BMMSCs transfected with Nc siRNA or HDAC9 siRNA, and those cells treated with CQ by western blotting. e, f The chromatin immunoprecipitation (ChIP) assay was performed to investigate whether HDAC9 could bind with the promoters of autophagy-related genes. Chromatin was isolated from young and aged BMMSCs (e) and aged BMMSCs transfected with Nc siRNA or HDAC9 siRNA (f), and incubated with HDAC9, acetylated-histone H3K9 (H3K9ac), and IgG antibodies. An IgG antibody was used as a negative control. Immunoprecipitation with specific HDAC9 or H3K9ac antibody. Data are presented as the mean ± SD of triplicate samples. *P < 0.05, **P < 0.01, ***P < 0.001. a, c One-way analysis of variance (ANOVA). e, f Unpaired two-tailed Student’s t test