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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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sIgM deficiency results in increased B cell receptor signaling in vivo. Representative flow cytometry plots and bar graphs show (a–c) the mean fluorescence intensity (MFI) for pSyk, pBtk levels and Nurr77-GFP expression in FO/T2 (blue), MZ (purple), CD21⁺ CD23⁻ (red), T1 (green) and NF (grey) B cells (as defined in Fig. S2) of sIgM ^+/+ or Nur77-GFP/sIgM ^+/+ (light colored bar) and sIgM ^−/− or Nur77-GFP/sIgM ^−/− (dark colored bar) mice. Data shown are from one representative experiment (a,b) or pooled (c) from two/three independent experiments. (d) Representative flow cytometry plots and dot plots show the frequencies of CD21⁺ CD23⁻ (red), FO (blue) and MZ (purple) B cells within B-2 cells of sIgM ^+/+ (●) and sIgM ^−/− (▼) mice treated with vehicle, and sIgM ^−/− (▲) mice treated with the Btk inhibitor Ibrutinib. All results show mean ± SEM, n = 4–6 mice per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (unpaired or paired t test or One-Way Anova test followed by Tukey’s test).