Figure 2.

Quantification and Limiting Dilution Analysis of Fluorescent Puncta after Subretinal Delivery of AAV Vectors

(A) Low and high magnification images of retinas at 24 h after subretinal injection of ∼0.25 μL of the indicated titers of AAV8-CMV-GFP. Scale bars, 50 μm. (B) Quantification of fluorescent puncta in cells of the retina with the conditions tested in (A). n = 477–1,733 cells from two to four animals per group. (C) Limiting dilution analysis of fluorescent puncta in retinas at 24 h p.i. based on the data shown in (B). Solid line depicts linear regression for the plotted data with 95% confidence interval shown in gray and goodness-of-fit measured by R². Dotted line depicts expected relationship if puncta count were to decrease by the square of the vector titer, as would occur if two independent AAV genomes in the same location were required to detect each punctum. Data shown are mean ± SEM. ∗∗p < 0.01, ∗∗∗∗p < 0.0001 by two-tailed Student’s t test with Bonferroni correction.