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. 2020 Mar 9;11(3):180. doi: 10.1038/s41419-020-2375-6

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© The Author(s) 2020

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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Fig. 6 — a Cdc20 and Cdh1 were knocked down in proliferative C2C12 using independent shRNAs (pLKO construct). Levels of Cdc20, Cdh1, FoxM1, and Tubulin (loading control) were determined by western blot analysis (n = 3). b SCs were isolated from FoxM1 deletion or overexpression mice and were cultured in vitro. Cdh1 proteins were immunoprecipitated from the cell lysates and analyzed by western blot (n = 3). WCL whole-cell lysates. c, d HA-ubiquitin (HA-Ub, ubiquitin expression construct with HA tag) plasmid was transfected with the indicated plasmids (FoxM1 overexpression plasmid, pCDH-FoxM1; Cdh1 knockdown plasmid, pLKO-shCdh1-1, pLKO-shCdh1-2) in proliferative C2C12 cells; 8 h after 10 μM MG132 treatment, HA-Ubiquitin (c) or FoxM1 (d) proteins were immunoprecipitated from cell lysates and analyzed by western blot (n = 4). WCL whole-cell lysates. e SCs were isolated from Cdh1^fl/fl and Cdh1^fl/flPax7-Cre (called Cdh1-cKO here) and were cultured in growth medium (activated state) or serum-free medium (reversed state) in vitro. Whole-cell lysates were subjected to western blot analysis using the indicated antibodies (n = 3). f The mononuclear cells were isolated from skeletal muscles of mice (2 months of age) and were stained with SC markers and FoxM1 fluorescent antibody. The intensity of FoxM1 was analyzed by flow cytometry. g The MFI analysis of FoxM1 in SCs (n = 3–4). Error bars represent the means ± SD. *p < 0.05; Student’s t test.