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. 2012 Mar 13;7(3):e33286. doi: 10.1371/journal.pone.0033286

Figure 1. AAV2/8 administration to newborn rats is associated with vector genome dilution.

Figure 1

A) Wild-type rats were injected at either postnatal day 4 (P4, upper pictures), or at P30 (lower picture) with 4×10e13 gc/kg of AAV2/8-TBG-eGFP. Animals injected at P4 were sacrificed at P15, P30 or P90, while those injected at P30 were analysed at P90. EGFP expression was confirmed under a fluorescence microscope on sections from transduced livers. Representative pictures from animals in each group are shown. Magnification: 20×. B) The eGFP expression levels were analyzed by Western blot of liver lysates from AAV-injected rats. The intensity of eGFP band was quantified, normalized on the corresponding tubulin band and expressed as fold of increase in each animal group compared to rats injected at P4 and sacrificed at P90. Results are reported as mean ± SE. The number of rats analyzed in each group is reported in each bar. *: p<0.05. The fold of increase of eGFP expression in livers from rats injected at P30 (white bar) compared to rats injected at P4 (light grey bar) and collected at P90 is 2.25±0.95. C) AAV vector genome copies per molecule of diploid genome (gc/mdg) were measured by Real-time PCR in Hirt DNA extracted from injected rats. Results are reported as mean ± SE. The number of rats analyzed in each group and statistical significance is reported as before.