Fig 2: Molecular mechanism of SHetA2: (A) Ishikawa cells were treated with SHetA2 (10 µM) for 4 h and co-immunoprecipitation was performed with mortalin antibody (Grp75)-tagged beads. Western blots (left panel) and densitometry analyses (right panel) was performed for IP3R, ALDH18A1, CTPS, MDH1, and ECHS1 to confirm the reduction of client protein co-immunoprecipitation. (B) Western blot analysis for the protein expression of IP3R, Cl. PARP, total caspase-3, P62, AIF, DNA damage marker (γH2AX) and LC3-II proteins in endometrial cancer cells treated with, or without, SHetA2 (10 µM) for 24 h. (C) Cell apoptosis was detected by Annexin-V/PI combined labeling with flow cytometry in endometrial cancer cells 24 h after treatment with SHetA2 (10 µM) or vehicle. (D) Caspase-3 activity assay for endometrial cancer cells treated with indicated dose of SHetA2 for 24 h. ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001 when compared with respective control.

Fig 3: AIF expression in RCC grades I, II and III and adjacent normal tissue. (A) IHC staining of AIF (brown) revealed a decrease in AIF expression with tumor grade. (B) Quantification of AIF expression in RCC grades I, II and III. AIF expression was significantly decreased in RCC tissues compared with that in adjacent normal tissues. *P<0.05, **P<0.01, ****P<0.0001. Scale bar, 400 µm. Normal, n=96; grade I, n=36; grade II, n=24; and grade II, n=36). AIF, apoptosis-inducing factor; RCC, renal cell carcinoma.

Fig 4: Immunolocalization of Cx43, ETFB and AIF proteins in freshly isolated cardiomyocytes from WT and Cx43KICx32 mice. (A) Confocal microscopy images showed a specific localization in cardiomyocyte membrane and mitochondria for Cx43; and presence in mitochondria for AIF and ETFB, in agreement with bibliographic information (upper right snapshots). Co‐localization (lower snapshots) of Cx43 with AIF or ETFB, and also AIF with ETFB, is quantitatively evaluated in (B) by analysis of composed images using 8–12 z‐stacks for each experimental condition, of at least three independent cardiomyocyte isolation events for each genotype. See text for comment on Manders' coefficients.

Fig 5: Morphology of the glomerular epithelial cells given anti-AIF antibodies. (a) P0 is the control, in which the mouse's kidney is not treated. (b) P1 is the kidney of mouse given cisplatin i.p on the 7th day. (c) P2 is the kidney of mouse injected with glutamine i.v 7 days in a row before being injected with cisplatin i.p on the 7th day. Positive if the cytoplasm and nucleus are colored brown.