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. 2012 Apr 9;209(4):775–792. doi: 10.1084/jem.20112422

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© 2012 Vilagos et al.

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Figure 3. — Role of EBF1 in late B lymphopoiesis. (A and B) Relative percentages and absolute numbers of B-1, MZ, and FO B cells were analyzed by flow cytometry of the spleen from Cd23-Cre Ebf1^fl/+ mice (gray bars) and Cd23-Cre Ebf1^fl/− littermates (black bars) at the age of 8 (A) and 16 (B) weeks. n, number of mice analyzed. Statistical data are shown with SEM and were analyzed by the Student’s t test. **, P < 0.01; ***, P < 0.001. (C) PCR genotyping of sorted FO, MZ, and B1 B cells of the indicated genotypes. The size (base pairs) and identity of the PCR fragments is shown to the left and right of the gel, respectively. C, genomic Ebf1^fl/fl or Ebf1^fl/Δ control DNA. (D) CD23⁺ splenic B cells of the indicated genotypes were isolated by MACS sorting before immunoblot analysis of serially diluted nuclear extracts (indicated by wedges) with EBF1, TBP, and Pax5 antibodies. The size (kilodaltons) of the proteins is indicated to the left. One of two immunoblots is shown. (E) Flow cytometric analysis of the cell surface phenotype of mature B cells (IgM^loIgD^hi) from the spleen of Cd23-Cre Ebf1^fl/+ mice (gray) and Cd23-Cre Ebf1^fl/− littermates (black line). (F) Analysis of MZ B cells as CD1d^hiCD21^hi splenic B cells in 8-mo-old Cd23-Cre Ebf1^fl/− and control Cd23-Cre Ebf1^fl/+ littermates.