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. 2018 Apr 24;9:1628. doi: 10.1038/s41467-018-04063-5

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© The Author(s) 2018

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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Fig. 3 — B cell-intrinsic Gal-3 deficiency favors ASC differentiation in vivo and in vitro and triggers spontaneous GC formation. a Schematic of the adoptive transfer experiment showing that B cells from Gal-3^+/+ WT and Gal-3 KO mice were injected iv into μMT mice. Bar graph showing the seric IgG2c and IgG3 concentration of recipient μMT mice at 2 weeks after cell transfer (μMT+Gal-3^+/+ B cells n = 5; μMT+Gal-3^−/− B cells n = 5). b Schematic of the adoptive transfer experiment showing that non-GC B cells from Gal-3^+/+ WT and Gal-3 KO mice were injected iv into CD45.1 WT mice. Bar graph showing the seric IgG2c and IgG3 concentrations of recipient WT mice at 2 weeks after cell transfer (CD45.1+Gal-3^+/+ B cells n = 5; CD45.1+Gal-3^−/− B cells n = 6). c IgG2c and IgG3 concentrations in the supernatants of B cells co-cultured with T cells (ratio 1:1) from the spleen of WT or Gal-3 KO mice and stimulated for 6 days with anti-CD3 and anti-IgM. The combinations of B and T cells are indicated (n = 4 for each condition). d Schematic of mixed BM chimeric mice containing a 80:20 ratio of μMT:Gal-3^+/+ WT or μMT:Gal-3 KO BM cells. e, f Representative plots and statistical analysis of the frequency of splenic GC B cells (e) and Tfh cells (f) from mixed BM chimeric mice shown in Fig. 3d (n = 7 for each group). g Frequency of splenic Ki-67⁺ B220⁺ B cells (left) and Ki-67⁺ CD4⁺ T cells (right) from mixed BM chimeric mice shown in Fig. 3d (n = 7 for each group). h Schematic of mixed BM chimeric mice containing a 70:30 ratio of CD45.1 Gal-3^+/+ WT:CD45.2 Gal-3 KO BM cells. i, j Representative plots and statistical analysis of the frequency of splenic GC B cells (i) and Tfh cells (j) from mixed BM chimeric mice shown in Fig. 3h (n = 9). k Frequency of splenic Ki-67⁺ B220⁺ B cells (left) and CD4⁺ T cells (right) from mixed BM chimeric mice shown in Fig. 3h (n = 9). Data are the mean ± s.e.m. *p < 0.05, **p < 0.005, ***p < 0.001 (unpaired (a–h) and paired (i–k) Student’s t-test). Data are representative of three (a–c) or two (d–k) independent experiments