Supplementary Figure 8: Monoallelic deletion of Irf5 abolishes the accumulation of ABCs and lupus development in DKO mice

(a,b) Percentage and numbers of B220⁺CD19⁺CD11c⁺T-bet⁺ (a) and of B220⁺CD19⁺CD21^–CD23^–CD11c⁺CD11b⁺ (b) B cells in the spleens of WT, Irf5^fl/fl DKO, Irf5^fl/– DKO, Cd11c-Cre Irf5^fl/– DKO and Cd21-Cre Irf5^fl/– DKO female mice (>20 weeks-old). Graphs show the frequencies and numbers for individual mice and mean value of 6 independent experiments (n = 7 WT, 5 Irf5^fl/fl DKO, 6 Irf5^fl/– DKO, 5 Cd11c-Cre Irf5^fl/– DKO and 6 Cd21-Cre Irf5^fl/– DKO). *** P = 0.0001; **** P < 0.0001. (One-way ANOVA followed by Bonferroni’s multiple comparisons test). (c) Flow cytometric analysis of CD11c⁺CD11b⁺ B cells in the spleens of WT, Irf5^fl/fl DKO and Cd11c-Cre Irf5^fl/– DKO female mice (>20 weeks-old). Representative FACS plots of 10 independent experiments is shown. (d) Spleen size of indicated mice. Graph shows numbers of individual mice and mean value of 10 independent experiments. (n = 9 WT, 10 Irf5^fl/fl DKO, 6 Irf5^fl/– DKO, 10 Cd11c-Cre Irf5^fl/– DKO and 5 Cd21-Cre Irf5^fl/– DKO female mice >20 weeks of age). * P = 0.0254; *** P = 0.0004; **** P < 0.0001. (One-way ANOVA followed by Bonferroni’s multiple comparison test). (e) Quantification of flow cytometric analysis of T_FH (CD4⁺CXCR5⁺PD1⁺Foxp3^–), GC (B220⁺FAS⁺GL-7⁺), and PC (B220^intCD138⁺) in spleens of WT, Irf5^fl/fl DKO, Irf5^fl/– DKO, Cd11c-Cre Irf5^fl/– DKO and Cd21-Cre Irf5^fl/– DKO female mice (>20 weeks). Graphs show percentages and numbers of specific cells types in individual mice and mean value of 10 independent experiments (n = 10 WT, 10 Irf5^fl/fl DKO, 6 Irf5^fl/– DKO, 10 Cd11c-Cre Irf5^fl/– DKO and 5 Cd21-Cre Irf5^fl/– DKO female mice). ** P < 0.01; *** P < 0.001; **** P < 0.0001. (One-way ANOVA followed by Bonferroni’s multiple comparisons test). (f) Quantification of flow cytometric analysis of T_reg (CD4⁺Foxp3⁺), activated T_reg (CD4⁺Foxp3⁺CD44⁺), and activated T cells (CD4⁺Foxp3^–CD44⁺) in spleens of WT, Irf5^fl/fl DKO, Irf5^fl/– DKO, Cd11c-Cre Irf5^fl/– DKO and Cd21-Cre Irf5^fl/– DKO female mice (>20 weeks). Graphs show frequencies and numbers of individual mice and mean value of 10 independent experiments (n = 9 WT, 10 Irf5^fl/fl DKO, 6 Irf5^fl/– DKO, 10 Cd11c-Cre Irf5^fl/– DKO and 5 Cd21-Cre Irf5^fl/– DKO female mice). ** P < 0.01; *** P < 0.001; **** P < 0.0001. (One-way ANOVA followed by Bonferroni’s multiple comparisons test).