Fig. 3

VO-OHpic could restore the normal mitochondrial morphology, suppress excessive ROS generation, and prevent the mitochondrial membrane potential (MMP) collapse induced by MPS. a Morphology of the mitochondria was stained with mito-tracker Green, and the representative fluorescence images were captured under a fluorescence microscope after treatment with 50-μM MPS or 50-μM MPS combined with 1-μM VO-OHpic for 48 h. b Morphology of the mitochondria was also observed and captured with a TEM after the same treatment. c Representative images of EPCs with intracellular ROS stained by the fluorescence probe DCFH-DA after treatment for 48 h. d Flow cytometric analysis of ROS production after staining with DCFH-DA. e Bar graphs showing the mean fluorescence intensity (MFI) of ROS levels in EPCs. Data are shown as means ± SD. f Representative fluorescence images of MMP after incubating with JC-1. Red fluorescence represents JC-1 aggregates in healthy mitochondria whereas green fluorescence represents JC-1 monomer indicating MMP dissipation. Merged images represent co-localization of the JC-1 aggregates and JC-1 monomers. g Representative graphs of the flow cytometric analysis after incubating with JC-1. FL1 represents JC-1 green and FL2 represents JC-1 red. h MMP were represented as green fluorescence ratio and data were presented as means ± SD. i MFF and FIS1 protein levels were determined by western blot analysis after treatment for 48 h. j Band density ratios of MFF and FIS1 to GAPDH in the western blots were quantified by densitometry. All experiments were repeated for three times; ∗P < 0.05 versus control, #P < 0.05 versus 50-μM MPS group. Error bar represents SD