Fig. 4

Reversible mitochondrial fragmentation and membrane potential loss in EtOH treated prRGCs. A Mito-tracker staining of mitochondria in normal prRGCs, EtOH (5%, vol/vol, 3 h) treated prRGCs, and prRGCs that were EtOH treated and then washed for 20 h (washed). Scale bar: 20 μm. B Cell viability was quantified by Hoechst/PI double staining. C Percentage of prRGCs with fragmented mitochondria. D and E The same prRGCs before EtOH treatment (Untreated), treated with 5% EtOH in culture medium for 1 h and 3 h, and then after washing EtOH and further culturing in fresh medium with (+) or without (−) BDNF and CNTF (Washed). Mitochondria were visualized by staining with TMRM. 25 cells were monitored. Scale bar: 20 μm. F Quantification of mean fluorescence intensity of TMRM staining. G-I Quantification analysis of mitochondrial number, mean mitochondrial volume, and total mitochondrial volume in individual prRGCs. Data are presented as the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. EtOH: ethanol