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. 2023 Mar 31;18(11):2526–2534. doi: 10.4103/1673-5374.371372

Figure 8.

Figure 8

Complementary modulation of Rheb and 4E-BP1 strikingly boosts RGC survival.

(A) AAV-EGFP, AAV-caRheb, AAV-caRheb + DN-S6K1, and AAV-caRheb + 4E-BP1-4A were administrated intravitreally to mice 2 weeks before ONC. Eyes were harvested on day 14 after ONC. (B) CaRheb + 4E-BP1-4A significantly increased the survival of RGCs while caRheb + DN-S6K1 significantly decreased RGC survival compared with caRheb alone. Fluorescent photomicrographs of retinal whole-mounts showed surviving Tuj1+ RGCs 14 dpc from eyes injected with AAV-EGFP, AAV-caRheb, AAV-caRheb + DN-S6K1, and AAV-caRheb + 4E-BP1-4A. Scale bars: 60 µm. (C) Quantification of surviving Tuj1+ RGCs from eyes injected with AAV-EGFP, AAV-caRheb, AAV-caRheb + DN-S6K1, and AAV-caRheb + 4E-BP1-4A. Data are presented as mean ± SD (n = 6–8). ****P < 0.0001 (one-way analysis of variance with Bonferroni’s post hoc test). AAV: Adeno-associated virus; caS6K1: constitutively active S6K1; DN-S6K1: dominate negative S6K1; dpc: days post crush; EGFP: enhanced green fluorescent protein; ONC: optic nerve crush; RGC: retinal ganglion cell; S6K1: mammalian target of rapamycin complex 1 downstream effector; Tuj1: β3-tubulin.