Figure 3.

SHARPIN is critical for germinal center B cell survival. (A,B) Flow cytometry analysis of apoptosis and necrosis (A) as well as caspase activation (B) in FAS^hiGL-7^hi GC B cells and FAS^–GL-7^– non-GC B cells in the spleen of B-Sharpin^+/+ or B-Sharpin^cpdmmice 14 d after immunization with NP-CGG/alum. (C) Flow cytometry analysis of CD4⁺ and CD8⁺ T cells (top panels) as well as CD4⁺CXCR5⁺PD-1⁺ Tfh cells (bottom) in mice, as in (A,B). (D) Gating strategy for flow cytometry analysis of Sca-1⁺c-Kit⁺Lin^– HSCs and the proportion of such cells in the bone marrow of Sharpin^+/+ and Sharpin^cpdm mice. (E) Flow cytometry analysis of (SSC^hiFSC^hi) granulocytes, (SSC^loFSC^lo) lymphocytes, and (SSC^loFSC^hi) monocytes in the spleen of CD45.1⁺ Sharpin^+/+/CD45.2⁺ Sharpin^cpdm mixed bone marrow chimera mice (mean and s.d., n=4). (F) Flow cytometry analysis of the proportions of CD45.1⁺ and CD45.2⁺ cells in different B cell subsets in the bone marrow and spleen of CD45.1⁺ Sharpin^+/+/CD45.2⁺ Sharpin^cpdm mice (mean and s.d., n=4). (G, H) Flow cytometry analysis of proportions of CD45.1⁺ and CD45.2⁺ cells in FAS^hiGL-7^hi GC B cells and FAS^–GL-7^– non-GC B cells (G) as well as survival and caspase 3 cleavage (H) in such cells in CD45.1⁺ Sharpin^+/+/CD45.2⁺ Sharpin^cpdm mice 14 d after NP-CGG/alum immunization (mean and s.d., n=4). * p<0.05; ** p<0.01; *** p<0.001; **** p<0.0001; t-test.