Supplementary Figure 3: Related to Fig. 3. Polyclonal T cells, but not plasma cells, induce inflammation in Ikzf1^B– mice.

a, Determination of the spleen weight and relative abundance of splenic plasmablasts (PB), effector/memory T, FO B and CD21^lo/–CD23^– B cells in experimental Ikzf1^B– Prdm1^B– mice (dark green, n = 8-10) and control Ikzf1^B+ Prdm1^B+ (gray, n = 7-8), Ikzf1^B– Prdm1^B+ (black, n = 12-14) and Ikzf1^B+ Prdm1^B– (light green, n = 7-9) mice at the age of 5-9 weeks. The frequency of the different cell types among total live splenocytes was determined by flow cytometry. b, Flow cytometric analysis of FO and CD21^lo/–CD23^– B cells (gated on B-2 cells), which were quantified in a. c, Relative frequency of splenic αβ T, NK, FO B and CD21^lo/–CD23^– B cells in experimental Eβ^–/– Ikzf1^B–mice (dark blue, n = 4-6) and control Eβ^+/– Ikzf1^B+ (gray, n = 5-8), Eβ^+/– Ikzf1^B– (black, n = 3-5) and Eβ^–/– Ikzf1^B+ (light blue, n = 10-13) mice at the age of 5-6 weeks. The frequency of the different cell types among total live splenocytes was determined by flow cytometry. d, Flow cytometric analysis of MZ B, CD21^lo/–CD23^– B, CD4⁺ T and CD8⁺ T cells from the spleen of J_HT mice that were reconstituted with bone marrow from Ikzf1^B+ mice (gray, n = 5 (untreated), 4 (anti-CD4), 3 (anti-CD8)) or Ikzf1^B– mice (black, n = 4 (untreated), 4 (anti-CD4), 4 (anti-CD8)), followed by intravenous injection of anti-CD4 or anti-CD8 antibodies at regular intervals (after day 1, week 1, 2, 3 and 4). The frequency of the different B and T cell types was determined 4.5 or 5 weeks after transplantation. e, Frequency of the indicated B and T cell types in the spleen of chimeric mice, which were generated by transplantation of Eβ^–/– Ikzf1^B– (dark blue) or Eβ^–/– Ikzf1^B+ (light blue) donor bone marrow into sub-lethally irradiated J_HT (n = 4-12), Rag2^–/– (n = 6-16) or OT-II TCR-tg Rag2^–/– (n = 2-5) host mice. The bone marrow chimeras were analyzed by flow cytometry 4-5 weeks post transplantation. Data in b are representative of mice analyzed in a, which were examined in 7 independent experiments. Data in c, d and e were analyzed in 5, 2 and 3 independent experiments, respectively. Statistical data (a,c,d,e) are shown as mean value with SEM and were analyzed by one-way ANOVA (Tukey; a,c,e), by multiple t-test (unpaired, two-tailed, Holm-Sidak; d) or by the Student’s t-test (unpaired, two-tailed; e (Eff/Mem T)); *P < 0.03, **P < 0.002, ***P < 0.0002, ****P < 0.0001. See Source Data for exact description of the mouse numbers (n) and P values. Each dot corresponds to one mouse.

Source data