FIG 5.

(A) rAAV5-, rAAV5+HS-, rAAV8(Y733F)-, and rAAV8(Y733F)+HS-mediated mCherry expression (red) following subretinal or intravitreal injection of between 2 × 10⁹ and 6 × 10⁹ vector genomes in C57BL/6J mice. The image of retina receiving AAV5 by subretinal injection was extended vertically to allow for visualization of transduction of RPE that was detached during the processing of tissue. IS/OS, inner segments/outer segments of photoreceptors; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer. Scale bar, 34 μm. (B) Transduction efficiency of unmodified AAV5 and AAV5+HS in 661W cone photoreceptor cells. Vectors were used at multiplicities of infection (MOI) of 10,000 and 50,000. mCherry expression was calculated with FACS by multiplying the percentage of positive cells by the mean fluorescence intensity in each sample. The red line indicates the value under which all signal is noise (no transduction is observed relative to that of blank cells).