Figure 1. BAFF-induced FM B cell... | Rockefeller University Press

$Figure 1. BAFF-induced FM B cell survival depends on MEK5 activity. (A and B) WT purified splenic FM B cells were cultured for 48 h with BAFF or control medium (–) plus 2 µM PD184352 (PD) or vehicle control (DMSO). (A) The fraction of live FM B cells was determined by flow cytometric analysis of 7AAD staining (mean ± SEM; pooled cells cultured in triplicates). Gating strategies used are shown in Fig. S1. (B) Lysates of purified splenic FM B cells were immunoblotted for the indicated proteins. (C and D) WT purified splenic FM B cells were cultured for 48 h with BAFF, α-IgM, or control medium (-), in the presence of 20 µM PD184352, 10 µM U0126, 10 µM BIX02188, 10 µM BIX02189, or control DMSO. The fraction of live FM B cells was determined by flow cytometric analysis of 7AAD staining (mean ± SEM; pooled cells cultured in triplicates). (E) Sorted splenic FM B cells (B220+AA4.1−IgM+CD23+) were cultured for 48 h with BAFF, α-IgM, or control medium (-) plus 5 µM BIX02188, 5 µM BIX02189, or control DMSO. The fraction of live cells was determined as in D. All results are representative of at least three independent experiments. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.$

Figure 1.

BAFF-induced FM B cell survival depends on MEK5 activity. (A and B) WT purified splenic FM B cells were cultured for 48 h with BAFF or control medium (–) plus 2 µM PD184352 (PD) or vehicle control (DMSO). (A) The fraction of live FM B cells was determined by flow cytometric analysis of 7AAD staining (mean ± SEM; pooled cells cultured in triplicates). Gating strategies used are shown in Fig. S1. (B) Lysates of purified splenic FM B cells were immunoblotted for the indicated proteins. (C and D) WT purified splenic FM B cells were cultured for 48 h with BAFF, α-IgM, or control medium (-), in the presence of 20 µM PD184352, 10 µM U0126, 10 µM BIX02188, 10 µM BIX02189, or control DMSO. The fraction of live FM B cells was determined by flow cytometric analysis of 7AAD staining (mean ± SEM; pooled cells cultured in triplicates). (E) Sorted splenic FM B cells (B220⁺AA4.1⁻IgM⁺CD23⁺) were cultured for 48 h with BAFF, α-IgM, or control medium (-) plus 5 µM BIX02188, 5 µM BIX02189, or control DMSO. The fraction of live cells was determined as in D. All results are representative of at least three independent experiments. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

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