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. 2020 Oct 26;10:18293. doi: 10.1038/s41598-020-75108-3

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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 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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Purified S. aureus MVs induce significant IFN-β mRNA expression in cultured murine macrophages. (A) Time-course analysis of RAW264.7 cells treated with MVs (5 µg/mL total protein) over a 24 h time period. Macrophages were homogenized at the indicated times points for RNA extraction and analyzed for β-actin and IFN-β mRNA expression using qPCR. The relative fold induction of a sample was calculated relative to the no treatment control sample using the Ct method. β-actin was used to normalize IFN-β mRNA expression in all samples. (B) Macrophages were stimulated with the indicated doses of S. aureus MVs for 3 h and the IFN-β mRNA expression relative to β-actin was determined by qPCR. (C) Macrophages were incubated with no treatment controls or with MVs (5 µg/mL) and IFN-α2, IFN-α4, or IFN-β mRNA expression was measured using qPCR. The data is represented as mean SEM, n = 3. Profiles are representative of at least two independent experiments. Statistical analysis was performed by one-way ANOVA with Tukey’s multiple comparison test (****p < 0.0001).