Fig. 2

prRGCs cannot survive from PS exposure after removing EtOH. A Live cell imaging of individual prRGCs without EtOH treatment (NC, negative control). Cells were imaged with annexin A5-FITC present in the culture medium for the duration of the experiment. Green (annexin A5-FITC) channel of the whole cell and bright field image of cell body were shown. Propidium Iodide (PI, red) was added and imaged at the last time point to detect cell viability. 12 cells were imaged in NC group. Scale bar: 50 μm. B Live cell imaging of individual prRGCs before EtOH (5%, vol/vol) treatment (untreated), during EtOH treatment (EtOH), and after washing away EtOH and further culturing cells in fresh culture medium for indicated time (washed). EtOH was removed when annexin A5-FITC staining was observed in the whole cell. Scale bar: 50 μm. C Live cell imaging of individual prRGCs before (untreated), during (EtOH), and after (washed) EtOH treatment. EtOH was removed when annexin A5-FITC staining was observed only in neurite. 45 cells were imaged. Scale bar: 50 μm. D Live cell imaging to monitor the degeneration of prRGCs neurites along with PS exposure. Scale bar: 50 μm. E The percentage of prRGCs with no PS, partial PS, or full PS. prRGCs were pre-treated with Z-VAD-FMK (zVAD, 20 μM) for 1 h, then co-treated with EtOH for another 8 h. 65-80 cells were counted in each group. BF: bright field. EtOH: ethanol. Time: hr.:min