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. 2020 Feb 13;10:2558. doi: 10.1038/s41598-020-59517-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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The effect of knockdown of COX-1 and COX-2 on Mbnl1 mRNA and MBNL1 protein expression in C2C12 cells. Western blotting (a,b,g,h) and real-time RT-PCR analysis (c–f) were performed using undifferentiated C2C12 cells or C2C12 cells differentiated for 5 days. The cells were treated with siRNA against COX-1 (siCOX1-a or siCOX1-b), COX-2 (siCOX2-a or siCOX2-b), both COX-1 and COX-2 (siCOX1a + 2a or siCOX1b + 2b), or control siRNA (siCont). Undifferentiated C2C12 cells were examined 2 days (mRNA levels) or 3 days (protein levels) after the transfection. Differentiated C2C12 cells were examined on day 5 of differentiation. Expression levels of COX-1, COX-2, Mbnl1 mRNA and MBNL1 protein are normalized to those of Gapdh and GAPDH, respectively, and also to siCont-treated cells. Mean and SD (n = 3 and 6 culture dishes for real-time RT-PCR and Western blotting, respectively) are indicated with individual values in red dots. *p < 0.05, **p < 0.01 and ***p < 0.001 by Student’s t-test with Bonferroni multiple comparison correction.