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. 2017 Nov 13;8:1462. doi: 10.1038/s41467-017-01602-4

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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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Fig. 7 — TACI requires mTOR to induce CSR in mouse MZ B cells. a FCM of p(S235/236)-S6 in splenic follicular and MZ B cells. Numbers in top panel indicate mean fluorescence intensity (MFI), whereas results from different experiments are summarized in bottom graph. Cells were gated as in Supplementary Fig. 11b. b ELISA of serum total and PCh-reactive IgM and IgG3 from WT and TNFSF13-Tg mice before (d0) and after (d7) daily i.p. treatment with rapamycin. c ELISA of serum TNP-specific IgM and IgG3 (d7) from WT and TNFSF13-Tg mice following i.p. immunization with TNP-Ficoll supplemented with daily i.p. injection of control vehicle or rapamycin. d ELISA of serum TNP-specific or total IgM and IgG3 (d7) from Mtor ^+/+ Cd21 ^cre/+ or Mtor ^fl/fl Cd21 ^cre/+ mice following i.p. immunization with TNP-Ficoll. e RT-PCRs of mRNAs for AID (Aicda), Iµ-Cµ, Iγ3-Cγ3, and GAPDH (Gapdh) in splenic resting B cells from Mtor ^fl/fl Cd21 ^cre/+ or Mtor ^+/+ Cd21 ^cre/+ mice treated for 3 days with APRIL in the presence or absence of LPS. Results are normalized to mRNA for Gapdh and quantified as relative expression (RE) compared to B cells from control Mtor ^+/+ Cd21 ^cre/+ mice. f ELISA of total IgG3 secreted by splenic resting CD43⁻ B cells from Mtor ^+/+ Cd21 ^cre/+ or Mtor ^fl/fl Cd21 ^cre/+ mice treated for 5 days with or without APRIL combined or not with LPS. g FCM of viable B220⁻CD138⁺ plasmablasts and PCs generated by splenic resting CD43⁻ B cells from Mtor ^+/+ Cd21 ^cre/+ or Mtor ^fl/fl Cd21 ^cre/+ mice following stimulation as in f. h qRT-PCRs of mRNAs for AID (Aicda), XBP-1 (Xbp1), and BLIMP-1 (Prdm1) in splenic resting CD43⁻ B cells from Mtor ^+/+ Cd21 ^cre/+ or Mtor ^fl/fl Cd21 ^cre/+ treated as in f. Results are normalized to mRNA for Gapdh and presented as RE compared to B cells from control Mtor ^fl/fl Cd21 ^cre/+ mice. i Frequency of viable splenic resting CD43⁻ B cells cultured as in f. Data summarize at least two experiments with at least two animals in each experimental group (a–g) and show one representative replicate (e, g). Error bars, s.e.m.; *p < 0.05, **p < 0.01, ***p < 0.001 (two-tailed Student’s t test)