Figure 2. c-FLIP controls Mo-MS viability.

(A) Diagram showing the role of c-FLIP in the inhibition of caspase 8 mediated apoptosis and necroptosis. (B) Flow cytometry analysis of BM-MS generated from FLIP^WT, RIPK3^WT, FADD^WT and c-FLIP^KO, RIPK3^KO, FADD^KO triple deficient mice. Numbers indicate the percentage of live cells (n = 3 independent experiments). (C) Flow cytometry analysis of BM-MS from Cflar^fl/fl; Rosa-CreERT2 mice treated with 4-OH tamoxifen at d3 of culture. Ethanol at the same final concentration as the 4-OH tamoxifen cultures served as the control. Numbers indicate the percentage of live cells (n = 3 independent experiments). (D) Cflar was exogenously deleted by 4-OH tamoxifen on d5 of culture using BM-MS from Cflar^fl/fl; Rosa-CreERT2 mice on a Ripk3^+/− or Ripk3^−/− background. Control cells received ethanol. On d6 Mo-MS were sorted and cultured with GM-CSF (50 ng/mL) for 24h, after which viability was assessed using V405 staining. (E) Cflar was exogenously deleted by 4-OH tamoxifen in the presence or absence of QVD (20 μM) on d5 of culture using BM-MS from Cflar^fl/fl; Rosa-CreERT2 mice. Control cells received ethanol and DMSO. On d6 Mo-MS were sorted and cultured with GM-CSF (50 ng/mL) in the presence or absence of QVD (20 μM) for 24h. Viability was assessed using V405 staining.