Fig. 6

Reversible mitochondrial fragmentation was accompanied by elevation of intracellular Ca²⁺ and occurred before cytochrome c release. A Live cell imaging of untreated and EtOH (5%, vol/vol, 3 h) treated prRGCs. Cells were stained with Mito-tracker (magenta) to show mitochondrial structure and Fluo-8 AM (green) to indicate intracellular Ca²⁺ level. Scale bar: 20 μm. B Quantification of intracellular Ca²⁺ mean fluorescence intensity in EtOH treated prRGCs. Data are presented as the mean ± SEM. **p < 0.01. C Immunofluorescence images of fixed prRGCs after EtOH treatment for 3 h. Mitochondria were stained by TOM20 antibody (magenta), cytochrome c by Cyto.c antibody (green), and nuclei by DAPI (blue). Scale bar: 20 μm. D Immunofluorescence images of fixed Hela cell after STS (500 nM) treatment for 2 h. Mitochondria were stained by TOM20 antibody (magenta) and cytochrome c by Cyto.c antibody (green). Scale bar: 10 μm