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. 2017 Sep 5;7:10490. doi: 10.1038/s41598-017-09981-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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Aggregation of IFNβ didn’t increase TLR2 and TLR4 response. (a,b) Betaseron and Rebif were aggregated by stirring as described in M & M. RAW-Blue cells and HEK-Blue hTLR2 and HEK Blue hTLR4 transfected cell lines were cultured for 24 hours in the presence or absence of aggregated and unaggregated samples. (c,d) Aggregation of Avonex was induced by oxidation as described in materials and methods. Both the oxidized and un-oxidized products were tested for immune activation using RAW-Blue cells. Immune activation is measured by quantifying NF-κB from the 24 h culture supernatant colorimeterically. Data is mean ± SD from the triplicate culture.