Figure 3.

Human RP2 AAV vector and its expression in Rp2-KO retina. (A) Schematic representation of the vector. (B) Immunoblot analysis using an RP2 antibody that recognizes both mouse and human RP2 proteins. The retinal lysate from an Rp2-KO mouse receiving subretinal administration of the AAV8-scRK-hRP2 vector for 1 month revealed a ∼40 kDa protein band corresponding to the human RP2 protein, which is similar to that detectable in human retinal lysate. The retinal lysate from a WT mouse revealed a band that migrates slightly faster than that from the human. β-Actin was used as loading controls. (C) Immunostaining of retinal sections from Rp2-KO mice 1 month after they received subretinal injection of vector or vehicle, using an antibody against both mouse and human RP2 proteins. A WT C57/Bl6 retinal section was used as positive control. Endogenous mouse RP2 protein was detected in multiple layers in WT retina, including the IS, OPL and IPL, which was not seen in the Rp2-KO retina. The vector-expressed human RP2 protein was primarily observed at the IS and nuclei of photoreceptors. Scale bar 50 μm. OS, outer segments; IS, inner segments; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer.