Fig. 2

O-GlcNAcylation in BAFFR signaling regulates the survival of mature B cells. a, b Frequency of Annexin V⁺ cells at various B-cell developmental stages, including pro-B (B220⁺ CD43⁺) and non-pro-B (B220⁺ CD43⁻) B cells (a), as well as late pre-B, immature and mature B cells within non-pro-B (B220⁺ CD43⁻) gate (b), in bone marrow of Ctrl and B-KO mice. c Frequency of BrdU⁺ mature B cells in Ctrl and B-KO bone marrow. d Frequency of Annexin V⁺ B220⁺ bone marrow and splenic B cells cultured for 72 h with (+) or without (−) recombinant BAFF (rBAFF, 200 ng ml⁻¹). Percentage of reduction of apoptosis in B220⁺ bone marrow and splenic B cells after BAFF treatment was calculated by (% of apoptosis in untreated group − % of apoptosis in rBAFF-treated group) / (% of apoptosis in untreated group). e Immunoblot of nuclear extracts isolated from rBAFF-treated Ctrl and B-KO splenic B cells at various time points with indicated antibodies against NF-κB subunits. Lamin B: nuclear protein loading control. f Immunoblot showing the activation of Erk5, p85/PI3K and Akt as well as Bim expression after rBAFF treatment in Ctrl and B-KO splenic B cells at various time points. Actin: protein loading control. The representative data from one of at least three independent experiments are shown. Quantification of results is shown in the right panel in a–d. The data are the mean ± s.e.m. (n = 3, a–d). *P < 0.05, **P < 0.01 (two-tailed unpaired t test). In e and f, the quantification of band intensity is indicated. N.S., not significant