Fig 2: Germinal Center B-cell responses to P. chabaudi infection are abrogated in Bcl6fl/flCD4-cre+/- mice, and reduced in SAP-deficient mice. Bcl6fl/flCD4-cre+/- and Sh2d1a-/- mice were infected with P. chabaudi via intra peritoneal injection of 105 iRBC, and the occurrence of GC B cells in the spleen was determined by flow cytometry at days 0, 15 and 35 post-infection, and by histology at days 0 and 35 post-infection. (a) Representative plots of naïve (top row) and infected (35 days post-infection, bottom row) Bcl6fl/flCD4-cre+/- and wt control mice. Gates show frequency of B220+ CD19+ IgDneg B cells expressing low levels of CD38 and high levels of GL-7. (b) Total numbers of GC B cells in Bcl6fl/flCD4-cre+/- and wt control mice. (c) Representative plots of naïve and infected Sh2d1a-/- and wt C57BL/6 control mice. (d) Total numbers of GC B cells in Sh2d1a-/- and wt C57BL/6 control mice. Bars represent median values. (e) Representative spleen sections of infected (d35) Sh2d1a-/- and wt C57BL/6 control mice; the white arrows point at individual examples of PNA+ GC structures. (f) Cumulative data displaying area, perimeter and number of GC structures determined by histology in spleen sections of infected (d35) Sh2d1a-/- and wt C57BL/6 control mice. Data are representative or pooled from at least two independent experiments and were obtained in groups of 3–8 mice per time point. Statistical significance was obtained using Student's t-test or Two-Way ANOVA with Tukey's multiple comparisons test. ND: Not done.

Fig 3: A poly(I:C)-induced maternal immune activation is not sufficient to drive Mll-AF4 leukemogenesis. (A) Survival curve of the progeny of pregnant dams injected with poly(I:C) at E14–E15 of embryonic development. (B) Body, (C) spleen, and (D) liver weights of mice at the end of study (18 months old). (E) Postmortem photos of sick mice 1463 and 1681 (564 and 516 days old) and control mouse 1684 (577 days old). Proportion of CD11b+ myeloid, CD3+ T-lymphoid, and B220+ B-lymphoid cells in the (F) bone marrow, (G) spleen, and (H) peripheral blood of sick mice 1463 and 1681. Peripheral blood counts of (I) white blood cells, (J) red blood cells, and (K) platelets of sick mice 1463 and 1681. (L) Postmortem photo of sick mouse NB1 (10 day-old) and age-matched control BL6/N mice. CD11b+ myeloid cells, CD3+ T-lymphoid cells, and B220+CD19+ and B220+CD19- B-lymphoid cells in the (M) bone marrow and (N) spleen of sick mouse NB1. (O) IgM expression on the B220+CD19– fraction of sick mouse NB1. (P) Overview of the hematopoietic features of all the sick mice (1463, 1681, and NB1). Statistical differences were assessed using a nonparametric Mann–Whitney U test with a bilateral p value or a Gehan–Breslow–Wilcoxon test (survival curve only) (*p < 0.05, **p < 0.01, ***p < 0.001).

Fig 4: ZC3H12D brings nex-IL1ß-mRNA into the nucleus.a Fluorescence confocal microscopy of ZC+RAW after application of 10 ng/mL non-labeled IL1ß-mRNA, FITC-labeled ßactin-mRNA, and IL1ß-mRNA with or without Cap-Poly(A) (CP). 3D images are shown (left). Quantitative analysis of RNA-FITC in the nucleus (right). IL1ß-stop-mRNA has three stop codon mutations in the CDS (top). The non-labeled IL1ß-mRNA was used as a basal control for the FITC signal (n = 5, 5, 5, 4, and 5 cells for non-labeled IL1ß-RNA, FITC-labeled ßactin-RNA, gapdh-RNA, IL1ß-RNA, and IL1ß-stop-RNA respectively). Repeated experiment data are shown in Supplementary Fig. S9. b System for the detection of external IL1ß-mRNA using human-mouse chimera IL1ß-mRNA (IL1ß-chimera) followed by reverse transcription with a 3'-mGSP and a 5'-human probe in the RT-qPCR analysis (top; more details in Supplementary Fig. S3b). Detection of IL1ß-chimeric-RNA entry into ZC+RAW. Nuclear RNA and cytoplasmic Hotair and ßactin normalized nuclear RNA and cytoplasmic RNA, respectively (n = 7, 8, 8, 8, and 8 biologically independent samples for 0, 30, and 120 min with IL1ß-chimera, 30 and 120 min with IL1ß-chimera-CP, respectively). c 3D images showing IL1ß-mRNA-FITC and IL1ß-stop-mRNA-FITC uptake in the nucleus of B220+CD11c+NK1.1+ cells derived from TCM-stimulated wild-type and Zc3h12d–/– mice 3 h after the application of 10 ng/mL RNA (top). Quantitative analysis of RNA-FITC in the nucleus of B220+CD11c+NK1.1+ cells from wild-type mice (bottom left, n = 6, 4, 6, 6, and 6 cells for non-labeled IL1ß-RNA, FITC-labeled ßactin-RNA, gapdh-RNA, IL1ß-RNA, and IL1ß-stop-RNA respectively) and from Zc3h12d–/– mice (bottom right, n = 6, 6, 6, 6, and 5 cells for non-labeled IL1ß-RNA, FITC-labeled ßactin-RNA, gapdh-RNA, IL1ß-RNA, and IL1ß-stop-RNA respectively). d Fluorescence confocal microscopy of ZC+RAW after the addition of FITC-labeled ßactin-mRNA, full length, CDS, or 3'-UTR of IL1ß-mRNA (top). Quantitative analysis of 10 ng/mL IL1ß-mRNA domain-dependent nucleus uptake (bottom; n = 10, 10, 11, 10, and 10 cells for non-labeled IL1ß-RNA, FITC-labeled ßactin-RNA, full length, CDS, and 3’UTR of IL1ß-RNA, respectively). In this analysis, 15-stacked 3D images were obtained from the bottom to the top of the nucleus. Bar, 5 µm. In the graphs, averages ± SEM and the results of one-way ANOVA with Bonferroni correction are shown. The P values are shown in the figure. Source data are provided as a Source Data file.

Fig 5: Administration of Mucin or Inulin Reduces Tumor Growth and Induces Anti-tumor Immunity(A) Growth of YUMM1.5 tumors that were subcutaneously transplanted in syngeneic C57BL/6 mice. Mice were provided with a control diet, 3% mucin in drinking water, or 15% inulin-supplemented chow starting 14 days before tumor inoculation (control, n = 12; mucin, n = 15; inulin, n = 15).(B) Quantification of tumor-infiltrating total CD45+ cells and effector (CD44hi) CD4+ or CD8+ T cells from mice treated as in (A) (control, n = 9; mucin, n = 10; inulin, n = 10).(C) Quantification of tumor-infiltrating, IFN-?-producing CD4+ T cells from mice treated as in (A) (n = 10).(D) Quantification of tumor-infiltrating total DCs and DC subsets in mice treated as in (A) (control, n = 9; mucin, n = 10; inulin, n = 10).(E) MFI of MHC class I and MHC class II on tumor-infiltrating DCs in mice treated as in (A) (control, n = 9; mucin, n = 10; inulin, n = 10).(F) Wild-type C57BL/6 mice (n = 12) were fed control or 15% inulin-supplemented chow starting 14 days before subcutaneous (s.c.) injection of YUMM1.5 melanoma cells (1 × 106). Anti-mouse Thy1.2 or control immunoglobulin G (IgG, 400 µg) were injected two times a week starting 3 days after tumor inoculation (n = 12). Tumor volume was assessed two times a week. FACS analysis revealed >90% depletion of blood CD4+ and CD8+ T cells on day 8 after tumor inoculation. Data are representative of three independent experiments (A–E) or one experiment (F). Graphs show the mean ± SEM. *p < 0.05, **p < 0.005, ***p < 0.001, ****p < 0.0001 by two-way ANOVA with Bonferroni’s correction (A and F) or by one-way ANOVA with Bonferroni’s correction (B–E).