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. 2020 Oct 22;11(10):891. doi: 10.1038/s41419-020-03112-6

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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. 1 — A miR-322/-503 was specifically expressed at the heart region and somites in E10.5 mouse embryos, which was determined by beta-galactosidase staining. White arrows pointed to the somites staining. B Expression patterns of miR-322 and miR-503 during normal myoblast differentiation. The expression levels of miR-322 and miR-503 were normalized to their day 0 values. C Inhibitors of miR-322 and miR-503 significantly repressed the expressions of myogenic markers (MyoD, MyoG, and Mef2C). All expression levels were normalized to the control group at day 0. D Inhibitors of miR-322 and miR-503 impaired myotube formation. Myotube formation was displayed by immunostaining of MF20 on day 6. E Inhibitors of miR-322 and miR-503 decreased myotube areas (left) and fusion index (right) of myoblast differentiation. F Overexpression of miR-322/-503 was verified by RT-qPCR. The expression levels of miR-322 and miR-503 were normalized to the control group. G miR-322/-503 overexpression significantly upregulated the expressions of myogenic markers (MyoD, MyoG, and Mef2C). All expression levels were normalized to the control group at day 0. H miR-322/-503 overexpression promoted myotube formation. Myotube formation was displayed by immunostaining of MF20 on day 6. I miR-322/-503 overexpression increased myotube areas (left) and fusion index (right) of myoblast differentiation. mirKD-Ctrl, C2C12/mirKD-Ctrl cells; mirKD-322, C2C12/mirKD-322 cells; mirKD-503, C2C12/mirKD-503 cells; Control, C2C12/control cells; miR-322/-503, C2C12/miR-322/-503 cell lines; * statistically significant (p < 0.05).