Fig. S6.

Tet2/3 DKO mice display abnormalities during B-cell development. (A) Representative flow cytometry analysis of splenocytes from the indicated genotypes. Numbers adjacent to outlined areas indicate % cells in each plot, and arrows indicate gating strategy Gates depict IgM⁺ B cells (B220⁺CD19⁺IgM⁺IgD^+/−), IgM⁻ B-cell progenitors (B220⁺CD19⁺IgM⁻IgD^–), immature IgM⁺ B cells (B220⁺CD19⁺IgM⁺AA4.1⁺), mature B cells (B220⁺CD19⁺IgM⁺AA4.1⁻), follicular (Fo) B cells (B220⁺CD19⁺IgM⁺AA4.1⁻CD1d^lo), and marginal zone (MZ) B cells (B220⁺CD19⁺IgM⁺AA4.1⁻CD1d^hi). (B) Graphs depict total cell numbers of myeloid cells (CD11b⁺) and IgM⁺ B cells (B220⁺CD19⁺IgM⁺IgD⁺) in the spleen (n = 3–7). (C) Representative flow cytometry analysis of splenocytes from SRBC immunized mice of the indicated genotypes. Numbers adjacent to outlined areas indicate % cells in each plot. Gates depict germinal center B cells [B220⁺CD19⁺CD95(Fas)⁺CD38^lo] and class switched IgG1⁺ B cells (B220⁺CD19⁺IgG1⁺CD38^lo). (D) Representative flow cytometry analysis of bone marrow cells from the indicated genotypes. Numbers adjacent to outlined areas indicate % cells in each plot, and arrows indicate gating strategy Gates depict total B cells (B220⁺CD19⁺), mature recirculating B cells (B220⁺CD19⁺IgM⁺AA4.1^–), immature IgM⁺ B cells (B220⁺CD19⁺IgM⁺AA4.1⁺), pro-B/pre-B cells (B220⁺CD19⁺IgM^–AA4.1⁺), pre-B cells (B220⁺CD19⁺IgM^–AA4.1⁺CD25⁺, ckit^–), and pro-B cells (B220⁺CD19⁺IgM^–AA4.1⁺CD25^–, ckit⁺). (E) Graph depicts the total cell number of mature recirculating B cells (B220⁺CD19⁺IgM⁺AA4.1^–) in the bone marrow (n = 3–7). All plots are gated on live singlets. Graphs depict mean values and SDs of the respective populations. Significance was calculated by the two-tailed Student t test (*P < 0.05; **P < 0.01; ***P < 0.001).