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. 2015 Sep 22;56(10):6049–6057. doi: 10.1167/iovs.15-17360

Figure 5.

Figure 5

(A) Flat mounts of GCL stained for TUNEL (green, white arrows). Compared to controls that were cultured for 24 hours (A, left panels), hypoxia/reoxygenation increased TUNEL-positive RGCs (A, middle panels). SNJ-1945 inhibited the increase in TUNEL-positive RGCs (A, right panels). Mounts countered stained for γ-synuclein (A, bottom panels) also demonstrate decreases in γ-synuclein in the RGCs during hypoxia/reoxygenation (middle panel), which was ameliorated by SNJ-1945 (right panel). Abbreviations as in Figure 2 legend. (B) Cross-sections show NFL/GCL stained for γ-synuclein. (C) Percentage of TUNEL-positive RGCs was calculated. Both hypoxia (H) and hypoxia plus reoxygenation (H&R) caused statistically significant increases in the percentage of TUNEL-positive RGCs and amelioration by SNJ-1945 (SNJ). Data are means ± SEM (n = 5). *P < 0.05 (Wilcoxon rank sum test). (D) Immunoblots for γ-synuclein from retinal explants cultured under hypoxia/reoxygenation conditions. (E) Band intensities from γ-synuclein in (D) normalized to GAPDH, confirming statistically significant decreases in γ-synuclein caused by hypoxia/reoxygenation and inhibition by SNJ-1945. Data are mean ± SEM (n = 9). *P < 0.05 relative to H&R. (F) Immunoblots for γ-synuclein from noncultured retinal soluble proteins incubated in the test tube without (−Ca) or with (+Ca) 10 mM Ca.