Figure 1.

Verification of Aipl1 expression. (A) Western blotting of 293T cells transfected with the plasmid construct and transduced by recombinant vector, AAV2-CMV-Aipl1 produces a ≈38 kDa protein band corresponding to Aipl1 protein. (B) Aipl1 immunofluorescence (green) is reduced in untreated Aipl1 h/h retina compared with treated retina which shows strong immunofluorescence localized mainly to the inner segments (IS) similar to that seen in wild-type retina. (C–E) Confocal images of double immunofluorescent microscopy for ß-PDE and Aipl1. In untreated retina, ß-PDE immunofluorescence (red) colocalizes with Aipl1 immunofluorescence (green) in the photoreceptor IS. Following subretinal injection of AAV2-CMV-Aipl1, ß-PDE immunofluorescence is translocated to the outer segments (OS) in the treated retina. Hence in treated retina, Aipl1 immunofluorescence in the IS is separate from ß-PDE immunofluorescence in the OS, which is similar to that found in wild-type retina. Cell nuclei are counterstained with Hoechst dye 33342 (blue). INL, inner nuclear layer; ONL, outer nuclear layer.