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. 2017 Aug 3;7:7219. doi: 10.1038/s41598-017-07575-0

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© The Author(s) 2017

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 siRNA-mediated TrxR1 inhibition reduces C2C12 cell growth and induces an early myogenesis-related gene expression pattern. C2C12 myoblasts were transfected with a scramble siRNA (scr siRNA) or a siRNA against TrxR1 sequence (TrxR1 siRNA) and cultivated in GM for 24–72 hours. (a) Western blot analysis showing the protein levels of TrxR1, Cyclin D1, p21 and Myf5 at 36 and 48 hours after transfection and the histogram relative to quantification of Cyclin D1 and Myf5 protein levels, represented as mean ± SEM of five different western blot as in panel a. The TrxR1/β-actin ratio of scr siRNA C2C12 is set = 1. (b) RT-qPCR analysis showing the mRNA expression levels of Myf5, Cyclin D1, p21 and Myogenin at 36 and 48 hours after transfection. Cyclophilin A mRNA was used to normalize the relative amount of each mRNA analysed. Results are represented as mean ± SEM of at least three different experiments. (c) MTS assay measured at 24, 48 and 72 hours after transfection. (d) Image (20x) showing cellular confluence at 72 hours. (e) Cytofluorimetric analysis showing cell cycle phase distribution.