Figure 3.

Transgenic BCLX protects lymphocytes more potently from apoptosis than BFL1. (A) Thymocytes from wild‐type, Vav‐BFL1 (both TG lines pooled), Vav‐BCLX (both TG lines pooled), Vav‐Mcl1 and Vav‐BCL2 TG mice were cultured in vitro for 72 h and apoptosis was assessed over time by flow cytometry. Cells negative for Annexin V and 7AAD were considered viable. (B) Thymocytes from wild‐type, Vav‐BFL1 (both lines pooled), Vav‐BCLX (both lines pooled), Vav‐Mcl1 and Vav‐BCL2 TG mice were either left untreated or exposed to 5 Gy γ‐irradiation, 100 nm staurosporine (STS) or 625 nm of the glucocorticoid (Glc), corticosterone respectively. Apoptosis was assessed after 20 h by flow cytometric analysis. (C) CD19⁺B220^loIgM⁻CD25⁺ pre‐B cells from the same mice were sorted by FACS from bone marrow and cultured for 48 h. Spontaneous apoptosis was assessed over time (left graph). In addition, sorted pre‐B cells were either left untreated or treated with 100 nm staurosporine (STS) or 625 nm corticosterone (Glc), respectively and apoptosis was assessed after 18 h by flow cytometry (right graph). (D) Gr‐1⁺ Granulocytes from mice of the indicated genotypes were sorted from the bone marrow and cultured for 48 h. Spontaneous apoptosis was assessed over time (left graph). Additionally, sorted granulocytes were cultured for 18 h either in the absence or presence of STS and apoptosis was assessed by flow cytometry (right graph). (E) Thymocytes were cultured in vitro for 8 h in the presence of 5 μm ABT‐737 and thereafter analysed for Annexin V and 7AAD positivity by flow cytometry. Statistical analysis was performed by using a two‐way ANOVA with Dunnett's multiple comparison. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; n ≥ 3 ± SD.