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Fig. 5 | Cell & Bioscience

Fig. 5

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. 5

NME6 localizes in mitochondria. a Exogenous NME6-GFP colocalizes with mitochondria in living cells. MDA-MB-231T cells transfected with pEGFPN1-NME6-186-GFP and pNME6-194-GFP were stained with MitoTracker™ (red) or cotransfected with pCFPmito to label mitochondria. Images were acquired 48 h post-transfection by confocal microscopy (scale bar: 10 µm). b Endogenous NME6 colocalize with mitochondria in fixed cells. HeLa cells transfected with pCFPmito to label mitochondria (cyan) were fixed and exposed to anti-NME6 antibody (green), before acquisition by confocal microscopy (scale bar: 10 µm). c Endogenous NME6 is enriched in Percoll-purified mitochondria. MDA-MB-231T cell fractionation was performed by differential centrifugation of cell homogenate (H) followed by a purification of crude mitochondria (MC) on Percoll gradient (C: cytosol, P: nucleus-rich pellet, MP: Percoll-purified mitochondria, CON: major contaminating band recovered after Percoll gradient). Fractions were evenly loaded on SDS-PAGE and analyzed by Western blot using antibodies against mitochondrial protein markers, markers for other compartments and NME6

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