(A) Schematic of the AAV construct for CRISPR-Cas9-mediated Spp1 knockdown in Kcng4-Cre-positive neurons (left) and timeline of experiment design for knockdown Spp1 at 2 weeks before SO injection and tissue harvest at 4 wpi (right).
(B) Control sgRNA/Cas9 Kcng4-YFP retina (top left), retina under SO treatment (top right), sgSPP1/Cas9-infected retina (bottom left), and sgSPP1/Cas9-infectedretina under SO treatment (bottom right), labeled with antibody to YFP.
(C) Quantification of normalized αRGC survival. n = 5 animals per condition.
(D) Schematic of the AAV construct for Spp1 overexpression in Foxp2-Cre-positive neurons (left) and timeline of experiment design for overexpressing Spp1 at 2 weeks before SO injection and tissue harvest at 4 wpi (right).
(E) Control AAV-expression retina (top left), retina under SOHU treatment (top right), AAV-Spp1-infected retina (bottom left), and AAV-Spp1-infected retina under SOHU treatment (bottom right), labeled with antibody to Foxp2.
(F) Quantification of normalized F-RGC survival.
Scale bars (B and E), 50 μm; n = 5 animals per condition. Paired t test; ns, not significant; **p < 0.01.