Figure 3

Depletion of splenic lymphoid and myeloid immune cells with the loss of MYSM1 DUB catalytic activity. Flow cytometry analyses were performed on (A,C–E) Mysm1^+/+, Mysm1^−/−, and Mysm1^DN/DN mice, and (B,F) Cre^ERT2 transgenic mice of Mysm1^fl/+, Mysm1^fl/fl, and Mysm1^fl/DN genotypes at > 20 weeks after tamoxifen treatment, to quantify (A,B) B cells (CD19⁺CD3⁻), CD4 T cells (CD3⁺CD4⁺CD8⁻), CD8 T cells (CD3⁺CD4⁻CD8⁺), NK cells (CD3⁻NK1.1⁺), and (E,F) monocytes (CD45⁺CD3⁻NK1.1⁻CD11b⁺Ly6C⁺Ly6G⁻), macrophages (CD45⁺CD3⁻NK1.1⁻CD11b⁺Ly6G⁻Ly6C⁻F4/80⁺CD64⁺), and neutrophils (CD45⁺CD3⁻NK1.1⁻CD11b⁺Ly6G⁺Ly6C⁻). The data is from (A,E) 3–10 mice per genotype consolidated from two independent experiments, or (B,F) 8–11 mice per genotype consolidated from three independent experiments. Bars represent means ± SEM; statistical analysis with ANOVA and Dunnett’s post-hoc test, comparing each group to the control, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, NS—not significant. (C,D) Representative flow cytometry plots of the spleen of Mysm1^+/+, Mysm1^−/−, and Mysm1^DN/DN mice gated on live cells and showing the depletion of (C) CD19⁺ B cells and (D) CD4⁺ and CD8⁺ T cells in Mysm1^−/− and Mysm1^DN/DN mice; the average frequencies of cells in the gates are presented as mean ± st. dev.