Figure 3. Ezh2 overexpression enhances RGC survival after ONC or excitotoxic injury.

(A) Representative immunofluorescence of retinal sections showing increased H3K27me3 levels in RGCs 2 weeks after intravitreal injection of AAV2-Ezh2, but not AAV2-Ezh2-Y726D. Retinal sections were stained with anti-H3K27me3 (green) and anti-Rbpms (red). Insets display enlarged images of RGCs in white, dashed boxes. Scale bars: 50 μm, 10 μm for enlarged images. (B) Quantification of fluorescence intensity of H3K27me3 immunoreactivity in RGCs in A (1-way ANOVA followed by Tukey’s multiple comparisons; P 0.0001; n = 3 mice for all; at least 150 RGCs from 10–12 nonadjacent sections were analyzed for each mouse). (C) Top: Experimental timeline. Bottom: Representative immunofluorescence of whole-mount retinas showing that overexpression of Ezh2 or Ezh2-Y726D improves RGC survival 2 weeks after optic nerve crush. Whole-mount retinas were stained with anti-Rbpms (green). Scale bar: 50 μm. (D) Quantification of RGC survival rate 2 weeks after ONC in C (1-way ANOVA followed by Tukey’s multiple comparisons; P = 0.0025; n = 3 mice for all; 6–9 fields were analyzed for each mouse). (E) Top: Experimental timeline. Bottom: Representative immunofluorescence of whole-mount retinas showing that overexpression of Ezh2 improves RGC survival 1 week after NMDA-induced excitotoxic injury. Whole-mount retinas were stained with anti-Rbpms (green). Scale bar: 50 μm. (F) Quantification of RGC survival rate 1 week after NMDA-induced excitotoxic injury in E (unpaired 2-tailed t test; P = 0.0042; n = 3 mice for both; 6–8 fields were analyzed for each mouse). **P 0.01, ****P 0.0001.