Fig. 4.

Combined loss of caspases-1/11/12 does not confer protection against treatment with a broad range of cytotoxic insults to cells in vitro. a Mouse dermal fibroblasts (MDF) from caspase-1/11/12 triple knockout or WT control mice were treated with thapsigargin (123 nM), taxol (10 μM), etoposide (300 μM), tunicamycin (1 μM), TNF (100 ng/mL) + SMAC mimetic (500 nM) and TNF (100 ng/mL) + SMAC mimetic (500 nM) + caspase inhibitor Q-VD-Oph (10 μM), and cell survival was measured at the indicated time points by flow cytometry. Graphs shown are for one representative experiment with n = 3 cell lines per genotype. Data are normalised to untreated control cells. Cell viability was measured by PI and Annexin V staining, followed by flow cytometric analysis. Data are presented as mean ± S.E.M. and were analysed using two-way ANOVA. b, c Primary BMDMs from caspase-1/11/12 triple knockout and WT control mice were treated with b TNF + SMAC mimetic + caspase inhibitor QVD-OPH (T + S + Q) to induce necroptosis or c TNF + SMAC mimetic (T + S) to induce apoptosis. Cell viability was measured at the indicated time points by Annexin V or Annexin V plus PI staining in the case of T + S + Q treatment, followed by flow cytometric analysis. All data were analysed using one-way ANOVA test. n ≥ 3. Data are presented as mean ± S.E.M.