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. 2021 Apr 19;12:658048. doi: 10.3389/fimmu.2021.658048

Figure 6.

Figure 6

SHARPIN inhibits IL-21-induced apoptosis in CD154-stimulated B cells. (A) Flow cytometry analysis of apoptosis (Annexin V+7–AADlo) and necrosis (Annexin V+7–AADhi) in freshly isolated Sharpin+/+ and Sharpincpdm B cells or after stimulation with CD154 (left) or αCD40 (right) plus nil or IL-21 for 96 h, with or without priming with CD154 or αCD40 for 4 h. (B, C) Flow cytometry analysis of apoptosis and necrosis in Sharpin+/+ and Sharpincpdm B cells after priming with TLR ligands (as indicated, B) or αIgD/dex (C) for 4 h and then stimulated with CD154 or αCD40 plus IL-21 for 92 h. (D) Flow cytometry analysis of the viability of Sharpin+/+ and Sharpincpdm B cells stimulated with CD154 plus IL-21 in the presence of nil or pan-caspase inhibitor Z-VAD-FMK, caspase 9-specific inhibitor Z-LEHD-FMK or caspase 8-specific inhibitor Z-IETD-FMK (all 20 μM). (E) Mitochondrial membrane potential in B cells stimulated with CD154 and IL-21 at indicated doses for 48 h (n=4, mean and s.e.m.; data at 4 units of CD154 were analyzed for statistical differences between Sharpin+/+ and Sharpincpdm B cells). *p < 0.05; **p<0.01; ***p < 0.001; t-test.