FIG. 2.

(A) Three-channel FACS analysis of live bone marrow (top two panels) and spleen B cells (bottom panel) prepared from 10- to 20-day-old pups carrying various E2A mutations. The genotype of each animal is indicated above each vertical panel, which is composed of three FACS analyses of the same animal. B220-FITC and 7AAD were included in all the stainings. In addition, CD19-PE or CD43-PE was used in the bone marrow samples, and IgM-PE was used in the spleen samples. The variation in the levels of CD19-PE staining is due to batch difference in the antibody used. To enrich lymphocytes and to exclude dead cells in the analysis, size gate and 7AAD gate were used for all the dot plots displayed. The relative percentages of pro-B, pre-B, and mature B cells in total live cells are given in each plot. FACS analysis of B cells in adult mice. Wild-type control, E47^bm homozygous and E47^bm/E12^ko transheterozygous mice were analyzed with B220 and 7AAD plus CD19 for bone marrow, IgM for spleen, and CD5 for peritoneum samples. Presentation of data is as described for panel A. (C) Average B-cell numbers as a percentage of total nucleated bone marrow or spleen populations. Both sample size and standard deviation are shown with each data bar. Four genotype groups of animals, wild-type (wt), E47^bm homozygous, E12^ko homozygous, and E47^bm/E12^ko transheterozygous, are included. Pup is equivalent to 10 to 20 days old; adult is more than 2 months old.