Caspase activation occurs after AIF translocation and nuclear condensation. Caspase activation occurs after AIF translocation and nuclear condensation.

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FIGURE S1 S1.c Scr. iPARG 30 nM H2O2 PAR iPARP-1 60 nM DAPI S1.a S1.b

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Caspase activation occurs after AIF translocation and nuclear condensation. Caspase activation occurs after AIF translocation and nuclear condensation. A, In wild-type (WT), but not PARP-1 KO, cortical cultures, exposure to NMDA (500 μm) results in the cleavage of caspase-3 revealed by immunoblot for the activated cleavage fragment. The pan-caspase inhibitor z-VAD.fmk (100 μm) prevents cleavage of pro-caspase-3. Wild-type cortical cultures were treated with 0.5 μm of staurosporine (S) for 8 hr as a positive control for activation of caspase-3. β-Tubulin is a loading control. These experiments were repeated at least two times with similar results. B, Time course of caspase activity monitored by cleavage of the fluorescent substrate by measuring the production of free AFC after NMDA (500 μm for 5 min) treatment. z-VAD.fmk blocks caspase activity induced by either NMDA or staurosporine (STS). These experiments were performed at least three times with similar results. Data are the mean values of three independent experiments ± SD. Significance was determined by a one-way ANOVA with Student's t test; *p < 0.05. Hongmin Wang et al. J. Neurosci. 2004;24:10963-10973 ©2004 by Society for Neuroscience