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. 2020 Feb 18;10:2845. doi: 10.1038/s41598-020-59444-y

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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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Reintroduction of StarD7 into the StarD7-KO C2C12 cells restored myogenic differentiation. (a) Cell lysates from WT + EV (empty vector), KO + EV, and KO + StarD7 C2C12 cells were separated by SDS-PAGE and analyzed by western blotting using anti-StarD7 antibody. GAPDH was used as a protein loading control. (b) WT + EV, KO + EV, and KO + StarD7 C2C12 cells were cultured in differentiation medium for 5 days, then immunostained with anti-MYH6 antibody (green). Nuclei were stained with DAPI (blue). Bars indicate 50 μm. (c) The fusion indexes of C2C12 cells were calculated, and are presented as the means ± S.D. ***P < 0.001 as compared with WT + EV cells, and ^†††P < 0.001 as compared with KO + EV cells (one-way ANOVA with Tukey’s post hoc test). (d) After inducing differentiation, the amounts of MYH4, MYH6, myogenin and myoD protein were analyzed by western blotting. Asterisk indicates a non-specific band. (e) The mRNA levels of MYH4, MYH6, myogenin, myomaker, myomerger (L), myomerger (S) and PGC-1α were quantified by qPCR after inducing differentiation. Data were normalized to the GAPDH level. Values shown are means ± S.D. from three independent culture dishes. *P < 0.05, **P < 0.01, and ***P < 0.001 as compared with WT + EV cells, and ^†P < 0.05, ^††P < 0.01, and ^†††P < 0.001 as compared with KO + EV cells (one-way ANOVA with Tukey’s post hoc test).