Live-dead assay of primary RGCs treated with endothelins.

<p><b>A.</b> Primary retinal ganglion cells were obtained from post-natal day 3–7 rat pups and seeded on coverslips. The cells were either untreated (control) or treated with 10 nM ET-1, 100 nM ET-1 or 100 nM ET-3 for 24 hrs. Following the treatments, a mixture of calcein AM (green fluorescence indicating living cells) and Ethidium homodimer (EtHD) (red fluorescence indicating dead cells) was added and cell viability assessed. <b>B.</b> Plot of percentage of dead cells after treatment of primary RGCs with ET-1 (10 nM), ET-1 (100 nM) and ET-3 (100 nM). The plot represents the average of three independent experiments and bars represent mean ± SEM. * indicates statistical significance (p<0.05) by Kruskal-Wallis One Way ANOVA on Ranks followed by Dunn's multiple comparison. Scale bar indicates 100 µm.