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. Author manuscript; available in PMC: 2012 Apr 5.
Published in final edited form as: Sci Signal. 2011 Apr 5;4(167):ra20. doi: 10.1126/scisignal.2000902

Figure 5. Pbm-AIF is resistant to release from mitochondria and subsequent nuclear translocation.

Figure 5

(A) Co-immunoprecipitation of PAR with WT-AIF-Flag and Pbm-AIF-Flag in post-nuclear fractions isolated from Hq cortical neurons 2 h after NMDA treatment (500 μM for 5 min), rabbit IgG (rIgG). (B) The signal was quantified and normalized to input. n = 5. (C) Subcellular localization of WT-AIF-Flag and Pbm-AIF-Flag in Hq cortical neurons 2 h after NMDA treatment, n = 4. (D) NMDA-induced AIF nuclear translocation in cortical neurons. DAPI, nuclei staining. Scale, 20 μm, n = 3. (E) Mitochondria isolated from cortical neurons were incubated with PAR for 30 min. The levels of AIF were determined in the mitochondria (left) and in the supernatant (right), n = 6. (F & G) WT-AIF-Flag or Pbm-AIF-Flag (500 ng/ml) were incubated with isolated mitochondria (1 mg/ml) in the absence or presence of PAR for 30 min. The supernatant was collected for analyses of AIF, Tom20, and Cyt C. AIF binding to mitochondria in the absence of PAR was regarded as 100% binding. ##p < 0.01, ###p < 0.001, compared to control (absence of PAR); * p < 0.05, **p < 0.01, ***p <0.001 by one-way ANOVA, n = 5.