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. 2009 Apr 20;185(2):291–303. doi: 10.1083/jcb.200811105

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© 2009 Manzl et al.

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Figure 5. — Caspase-2 processing occurs in thymocytes lacking the PIDDosome. (A) Thymocytes isolated from wild-type (wt) and pidd^−/− mice were cultured without further treatment or stimulated with etoposide. Caspase-2 processing and cleavage of effector caspase-3 was monitored over time by Western blot analysis. The asterisk indicates a residual caspase-2 signal remaining upon reprobing with anti-GAPDH antibody. (B and C) Caspase-2 and -3 processing in etoposide-treated (B) or γ-irradiated thymocytes (C) of the indicated genotypes. tg, transgenic. (D) Thymocytes isolated from wild-type, pidd^−/−, puma^−/−, and pidd^−/−puma^−/− mice were cultured without further treatment or after exposure to γ irradiation for 6 h. Caspase-2 processing and cleavage of effector caspase-3 were monitored by Western blot analysis. After detection of caspase-2, membranes were first stripped and reprobed with anti–active caspase-3 antibody and finally probed without stripping with an antibody recognizing GAPDH or α-tubulin to compare protein loading. DKO, double knockout.