Figure 1 | Cell Death & Disease

Figure 1

From: Death of p53-defective cells triggered by forced mitotic entry in the presence of DNA damage is not uniquely dependent on Caspase-2 or the PIDDosome

Figure 1

Chk1 inhibition fails to sensitize p53-deficient lymphocytes or MEF to DNA damage. Primary thymocytes (a), stimulated T- (c) or B-cell blasts (d) were treated with Gö6976 and/or exposed to IR or both. As a control, cells were treated with staurosporine (STS). Cell viability was assessed after 24 h using AnnexinV/PI staining and flow cytometric analysis. To account for spontaneous cell death of lymphocytes in culture, specific cell death was calculated throughout using the equation (induced apoptosis−spontaneous cell death)/(100−spontaneous cell death). In parallel, thymocytes were harvested and protein lysates were separated by SDS-PAGE to monitor cleavage of Caspase-2 and Caspase-3. Membranes were reprobed with an anti-GAPDH antibody to control protein loading (b). Viability of primary (e) and SV40 immortalized MEFs (f) derived from E14.5 embryos of the indicated genotypes were assessed using sub-G1 staining and flow cytometric analysis. Bars represent means±S.E. of ≥3 independent experiments per genotype and treatment. *Indicates significant differences between wt or Casp2−/− cells and p53−/− or DKO cells (P<0.05) after IR or Gö+IR treatment. No significant differences were noted in (a, c and d) between Gö and Gö+IR treatment

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