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Fig. 7 | Cell & Bioscience

Fig. 7

From: NME6 is a phosphotransfer-inactive, monomeric NME/NDPK family member and functions in complexes at the interface of mitochondrial inner membrane and matrix

Fig. 7

NME6 overexpression affects cellular distribution and ADP-stimulated respiration of mitochondria, but not their membrane potential. a Confocal images of MDA-MB-231T cells stained with Mitotracker Green, either wild-type (Ctrl) or clones stably overexpressing NME6-186-FLAG about 10-times (KI-186-7) or about 20-times (KI-186-18 and -26) as compared to endogenous NME6 in Ctrl (scale bar: 10 µm). 3D animations of all shown cells are available as Additional files 8, 9, 10, 11: Movie S4, S5, S6, S7. Note the altered distribution of mitochondria in particular in KI-186-18 and -26 cells, forming dense clusters around the nuclei. b–e Oxygraphy analysis of respiration in the cells shown in (a) that were digitonin-permeabilized and supplied with substrate (LEAK), stimulated with ADP (OXPHOS), and tested for mitochondrial membrane integrity (addition of cytochrome c). b Cellular oxygen consumption with glutamate/malate. c OXPHOS/LEAK ratios for b. d Cellular oxygen consumption with succinate. e OXPHOS/LEAK ratios for d. f Mitochondrial mass determined by Mitotracker Green staining. g Mitochondrial membrane potential determined by TMRM and corrected for mitochondrial mass (for details see “Material and methods”). All data are given as mean ± SEM (n = 3). For comparison between Ctrl and NME6 overexpressing clones, significance is given as **p < 0.01; *p < 0.05 (Student’s test)

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