Fig. 1: EPC proliferation inhibition and mitochondrial dysfunction in atherosclerotic mice.
a ApoE−/− mice were fed with HFD and CCK-8 was employed to measure EPC proliferation. CCK-8 results showed that EPC proliferation decreased 25.24% at 8 weeks and 47.31% at 16 weeks respectively in comparison to that of ND. b EPCs from ND and atherosclerotic mice were seeded on E-plates and the proliferation ability was real-timely recorded. The normalized cell index indicated EPC proliferation from atherosclerotic mice decreased compared with those from ND mice. Representative graphs were shown from 3 independent experiments. c Mitochondrial membrane potential was measured by JC-1. Quantitative analysis of red and green fluorescence showed that the ratio of red related to green fluorescence decreased to 1.204 in HFD8w group (p < 0.01) and to 0.458 in HFD16w group (p < 0.01). d Mitochondrial ROS production was evaluated by MitoSOX. Quantitative analysis indicated that red fluorescence increased in HFD8w group (p < 0.01) and this was more pronounced in HFD16w group (p < 0.01). e EPCs were incubated with JC-1, a fluorescent dye of mitochondrial membrane, for 25 min. Fluorescence was visualized via LCSM. Representative images showed the fluorescence intensity in different groups, scale bar: 50 μm. f Mitochondrial superoxide levels were labeled with MitoSOX fluorescence indicator for 20 min. Fluorescence was visualized via LCSM. Representative images showed the fluorescence intensity in different groups, scale bar: 50 μm. g Mitochondrial morphology was captured by transmission electron microscope. Representative images showed the mitochondrial morphology in different groups, scale bar: 1 μm. (Cells were isolated from 3 mice for 1 experiment and 5 independent experiments were performed, mean ± SD, **P < 0.01).
