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. 2017 Nov 7;7:14641. doi: 10.1038/s41598-017-15269-w

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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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Differentiation of C2C12 cells into skeletal myotubes enhances CHIKV RNA replication. C2C12 cells were differentiated via serum starvation. (a) Bright field image showing the different morphology of differentiated cells compared to C2C12 cells maintained in complete culture medium (undifferentiated cells). Bright field images were obtained on an IncuCyte ZOOM instrument equipped with a 10 × objective. (b) Western blot for skeletal myosin, a marker of skeletal muscle tissue, is highly expressed in differentiated cells (D) compared to undifferentiated (U). (c) Both differentiated and undifferentiated C2C12 cells were transfected with CHIKV-D-Luc-SGR RNA. Cells were lysed at indicated time points and assayed for luciferase, data analysed by parametric T-test (**p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001). (d) Confocal images of undifferentiated (U) and differentiated (D) cells transfected with nsP3-mCherry/SG-GLuc.