FIGURE 1.
Flow-cytometric analysis of cell surface Ig expression and B cell development in spleen and bone marrow. A, FACS analysis of cell surface Ig expression in WT and Sis−/− mice. Single cell suspensions from spleen were simultaneously stained with anti-B220 and anti-Igκ antibodies (upper); or anti-B220 and anti-Igλ antibodies (lower). Stained cells were analyzed by FACS. Only cells residing in the lymphocyte gate were analyzed. Percentages of cells residing in various windows are shown in the figure sub-panels. Data are representative of independent FACS analyses from at least 6 mice of each genotype. B, FACS analysis of transitional, follicular mature, and marginal zone B cells in WT and Sis−/− mice. Single cell suspensions from spleen were simultaneously stained with anti-IgM and anti-IgD antibodies (upper); or anti-CD21 and anti-CD23 antibodies (lower). C, FACS analysis of cell surface Ig expression in bone marrow of WT and Sis−/− mice. Single cell suspensions from bone marrow were stained and analyzed as described in A. Only cells residing in the lymphocyte gate were analyzed. Data are representative of independent FACS analyses from at least 6 mice of each genotype. D, FACS analysis of B cell development in bone marrow of WT and Sis−/− mice. Single cell suspensions from bone marrow were stained with anti-B220 and anti-c-kit antibodies to assay for pro-B cells (upper), anti-B220 and anti-CD25 antibodies to assay for pre-B cells (middle), or anti-B220 and anti-IgM to assay for immature and mature B cells (lower).