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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. 1 — MZ B cells couple increased TACI expression with enhanced mTORC1 signaling. a Transmission electron microscopy of human splenic follicular (FO) and MZ B cells. Original magnification ×13,500 with 6× enlargement. Scale bars, 1 or 0.2 μm. b Flow cytometric analysis of LysoTracker, ER-Tracker, MitoTracker, and Golgi in human splenic FO and MZ B cells. SSC-A side scatter-area. c, d Transcriptome analysis of human splenic FO and MZ B cells. The heat map shows genes upregulated in FO (1302) and MZ (800) B cells and highlights a core MZ B-cell gene signature. The volcano plot summarizes comparably and differentially expressed genes. Red high expression, blue low expression, FC fold change, FDR false discovery rate. e FCM of human splenic FO and MZ B cells undergoing proliferation-induced CFSE dilution or CD19⁺CD27^hiCD38^hi plasmablast differentiation following incubation with or without APRIL and/or CpG for 5 days. f Gene ontology analysis showing top seven biological processes upregulated (red) and downregulated (blue) by human MZ B cells compared to FO B cells. Numbers indicate genes. g, h Heat maps list top 10 genes similarly enriched or depleted in GSEA comparing genes expressed by human splenic MZ and FO B cells linked to mTORC1 activation (HALLMARK_MTORC1) (g) or PC differentiation (GSE22886) (h). i IFA of human spleen stained for IgD (red), p(S235/S236)-S6 (green), and MAdCAM-1 (blue). Scale bars, 100 μm (main image) or 10 μm (inset). j, k FCM of p(T308)-AKT, p(S473)-AKT, p(S235/S236)-S6, and CD98 in human FO and MZ B cells. Cells were gated as in Supplementary Fig. 11a (b, j, k). Data show one representative experiment of at least three with similar results (a, b, i–k), depict at least three biological replicates for each cell type (c, d, f-h), or summarize at least two experiments with at least two donors from each experimental group (e). Error bars, s.e.m.; *p < 0.05, **p < 0.01, ***p < 0.001 (two-tailed Student’s t test)