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Figure 7 | BMC Cancer

Figure 7

From: Annonaceous acetogenin mimic AA005 induces cancer cell death via apoptosis inducing factor through a caspase-3-independent mechanism

Figure 7

RIP1 is required for AA005-induced cell death. (A) Immunoblotting analysis of the expressional level of RIP-1 after 1 μM AA005 treatment or 8 h MNNG treatment for the indicated times, standardized to actin. (B) Flow cytometry analysis of AA005 or MNNG induced cell death in the presence of RIP-1 inhibitor Necrostatin-1 (Nec-1; 100 μM). Numbers are mean values of three independent experiments ± S.D. *P < 0.05 versus AA005- or MNNG-treated group). (C) Immunoblotting analysis of AIF translocation by subcellular fractionation in the presence of either RIP-1 inhibitor Necrostatin-1 (Nec-1; 100 μM) or PARP inhibitor Olaparib (AZD2281; 50 μM). Lamin B and COX IV were used as nuclear and mitochondrial marker proteins, respectively. (D) SW620 cells were transfected with retrovirus harboring either scrambled negative control (NC) or siRNAs against AIF (designated as A3 and A5). AIF and RIP-1 were examined by western blots, standardized to actin. Experiments in (A), (C) and (D) were repeated at least three times with similar results.

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