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. 2024 Feb 28;221(4):e20221080. doi: 10.1084/jem.20221080

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© 2024 Zhang et al.

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Figure 3. — B cell phenotypes in the spleens of Sh2b3^Δ, Sh2b3^E372K, and Sh2b3^R530Q mice, and peripheral blood of SLE patients carrying SH2B3 variants R566Q and Q540X. (A) Representative flow cytometric plot showing the gating of mature (B220⁺CD93⁻) and transitional (B220⁺CD93⁺) B cells. (B) Dot plots showing the percentages of splenic transitional B cells in Sh2b3^Δ (left), Sh2b3^E372K (middle), and Sh2b3^R530Q (right) mice. (C) Ratios of CD45.2⁺/CD45.1⁺ transitional B cell percentages in the spleens of 50:50 BM chimeras from CD45.1-Sh2b3^+/+ and CD45.2-Sh2b3^+/+ (n = 10) or Sh2b3^E372K/E372K (n = 10) mice. Student’s t test was used for statistical analysis. ####: P < 0.0001. Results are representative of two independent experiments. (D) Representative flow cytometric plot showing the gating of T1 (IgM⁺CD23⁻), T2 (IgM⁺CD23⁺) and T3 (IgM⁻CD23⁺) B cells. (E) Dot plots showing the percentages of splenic T1 B cells in Sh2b3^Δ (left), Sh2b3^E372K (middle), and Sh2b3^R530Q (right) mice. (F) Percentages of human transitional B cells in the peripheral blood of HCs (n = 77), SLE patients carrying SH2B3 variants (R566Q in turquoise, n = 2, and Q540X in purple, n = 1) and other SLE patients (n = 23). (G) Representative flow cytometric plot showing the gating of CD21⁻CD23⁻ mature B cells and MZ B cells (CD21/35⁻CD23⁺). (H) Dot plots showing the frequencies of MZ B cells as percentages of total splenic B cells in Sh2b3^Δ (left), Sh2b3^E372K (middle), and Sh2b3^R530Q (right) mice. (I) Frequencies of CD21⁻CD23⁻ mature B cells as percentages of total splenic B cells in Sh2b3^Δ (left), Sh2b3^E372K (middle), and Sh2b3^R530Q (right) mice. (J) Percentages of human CD21^lo/− B cells in the peripheral blood of HCs (n = 77), SLE patients carrying SH2B3 variants (R566Q in turquoise, n = 2, and Q540X in purple, n = 1), and other SLE patients (n = 23). For panels B, E, H, and I, sample numbers are listed as follows: Sh2b3^Δ panels: +/+ (n = 5), Δ/+ (n = 7 in B and E, n = 9 in H and I), Δ/Δ (n = 6 in B and E, n = 9 in H and I); Sh2b3^E372K panels: +/+ (n = 4), E372K/+ (n = 3), E372K/E372K (n = 4); Sh2b3^R530Q panels: +/+ (n = 5), R530Q/+ (n = 7), R530Q/R530Q (n = 6). Each dot represents one mouse, and means are shown as bars. Results are representative of three independent experiments. One-way ANOVA with multiple comparison and Dunnett’s correction was used for statistical analysis. Significance levels are indicated with asterisks. For panels F and J: Brown–Forsythe and Welch ANOVA tests were used for statistical analysis. Dollar signs are used to indicate significance levels. Significance level criteria are described as follows: */$: P < 0.05, **: P < 0.01, ***: P < 0.001, ****/####/$$$$: P < 0.0001.