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. 2017 Jun 14;7:3540. doi: 10.1038/s41598-017-03688-8

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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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Antigen-specific secreted IgM limit self-antigen induced B cell receptor signaling. (a) Quantification of hen egg-white lyoszyme (HEL) specific IgM in the plasma of MD4, RAG1^−/− and wild-type (WT) mice by ELISA. (b) Representative flow cytometry plots and bar graphs represent the mean fluorescence intensity (MFI) for pBtk in purified B-2 (B220⁺ CD43⁻) cells from MD4^+/− mice stimulated with HEL for 3 minutes in presence of either WT (grey), RAG1^−/− (purple) or MD4 plasma (blue). ELISA quantification of (c) HEL-specific and (d) total IgM in MD4 plasma treated with either unconjugated or BSA- or HEL-conjugated microspheres. (e) Representative flow cytometry plots and bar graphs show the MFI for pBtk in purified B-2 cells from MD4^+/− mice stimulated with HEL for 3 minutes in the presence of plasma from MD4 mice that has been treated with either unconjugated or BSA- or HEL-conjugated microspheres. All results show mean ± SEM, ****P < 0.0001 (unpaired t test or One-Way Anova followed by Tukey’s test). n.d.: not detechtable.