Intravitreal injection of AAV-hSyn-SARM1.DN-EGFP drives expression in RGCs but not oligodendrocytes. (A) Representative confocal single section images of retina (cross-section) following intravitreal injection of AAV-hSyn-SARM1.DN.EGFP. Retina are stained for nuclei (DAPI, blue), axons (TUJ1, yellow), RGC nuclei (Brn3a, red), and SARM1.DN (green). The white line is the region of interest for the analysis of pixel intensity of each channel. Scale bar, 20 µm. (B) Pixel intensity analysis for the region of interest from A. The RGC nuclear marker Brn3a overlaps with DAPI staining (arrowhead) while the adjacent GFP signal of SARM1.DN overlaps with TUJ1 (arrow). SARM1.DN is detected in the cytosol of RGCs. (C) Representative confocal single section images of optic nerve (longitudinal section) following intravitreal injection of AAV-hSyn-SARM1.DN.EGFP. Optic nerve is stained for nuclei (DAPI, blue), axons (TUJ1, yellow), oligodendrocyte nuclei (CC1, red), and SARM1.DN (green). The white line is the region of interest for the analysis of pixel intensity of each channel. Scale bar, 20 µm. (D) Pixel intensity analysis for the region of interest from A. The mature oligodendrocyte nuclear marker CC1 overlaps with DAPI staining (arrowhead) but is distant from the axonal staining of SARM1.DN and TUJ1 (arrow) consistent with RGC expression of SARM1.DN. AU, arbitray units; ROI, region of interest.