Figure 2.

Colabeling of EGFP transduced cells with cell markers in the inner retina

Adult mice received 5.0 × 10¹⁰ vg/animal of AAV-PHP.eB-CMV-EGFP via TV injection or 7.5 × 10⁸ vg/eye of AAV-PHP.eB-CMV-EGFP or AAV2/2-CMV-EGFP (control) IVT (n = 3–4). Eyes were enucleated, fixed in 4% PFA, cryosectioned, and stained with immunohistochemistry for retinal cell markers at 1 month post-delivery. (A–F) GCL. (A–C) EGFP fluorescence (green), (D–F) overlay of EGFP fluorescence (green), RBPMS immunohistochemistry (retinal ganglion cell marker, magenta), and nuclear counterstain (DAPI, blue). Arrowheads indicate examples of EGFP⁺ retinal ganglion cells. (G–R) INL. (G–I) EGFP fluorescence (green), (J–L) overlay of EGFP fluorescence (green), VSX2 immunohistochemistry (bipolar cell marker; magenta), and PAX6 immunohistochemistry (amacrine cell marker; light blue). Upward arrowheads indicate examples of EGFP⁺ bipolar cells; downward arrowheads indicate examples of EGFP⁺ amacrine cells, while arrows indicate examples of EGFP⁺ horizontal cells. (M–O) EGFP fluorescence (green). (P–R) Overlay of EGFP fluorescence (green) and CRALBP immunohistochemistry (Müller cell marker; magenta). Arrows indicate horizontal cells, while triple arrowheads mark the layer of Müller cell bodies. Scale bars represent 50 μm (C, I, and O).