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. 2021 Feb 10;11:3476. doi: 10.1038/s41598-021-82706-2

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© The Author(s) 2021

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 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/.

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Changes in the expression profile of miRNAs in human skeletal myocytes with MEF2C depletion. (a) MA-plot showing the prediction of differentially expressed miRNAs between cells treated with control and MEF2C-specific siRNAs. The red solid dots represent the statistically significant results for miRNAs downregulated in cells treated with siMEF2C_1; the green solid dots represent significantly upregulated miRNAs by siMEF2C_1. Empty dots represent miRNAs deregulated by both siMEF2Cs (_1 and_2). The experiment was performed in HSkM cells in triplicates. (b) Validation of the results from (a) in HSkM with MEF2C KD. The expression of 14 mature miRNAs was quantified using polyA-RT-qPCR analysis. The expression level of miRNAs were normalized to the level of the small nuclear U6RNA. Green/dark green and red/orange bars represent miRNAs predicted to be upregulated or downregulated according to RNA-seq after treatment with siMEF2C_1 or siMEF2C_2, respectively. The results are average values from three independent experiments + / − SD (* P < 0.05). (c) Relative expression changes of pri-miRNAs quantified by RT-qPCR assays divided into two groups (miRNAs predicted to be upregulated or downregulated according to RNA-seq results); all data for pri-miRNA were averages from three independent experiments + / − SD normalized to mRNA of GAPDH (* P < 0.05). MiRNAs from the same genetic cluster are marked with a blue line. (d) Expression changes of miRNAs coming from the same pre-miRNA hairpin or from the same genetic cluster (marked with blue lines) based on RNA-seq data. MiRNA candidates which could be regulated transcriptionally or posttranscriptionally are indicated.