FIG 2.

The BR is not the determinant for serotype specificity of AAV capsid assembly. (A) Schematic of different AAP mutants. The T/S of AAP1 was replaced by EGFP (AAP1E). The BR was replaced with those of other serotypes, including AAP4 (4BR), AAP5 (5BR), and AAP9 (9BR). (B and C) Representative Western blot images of VP (B) and AAP (C) expression as described in Fig. 1B. (D) Confocal microscopy of different AAP1E derivatives. AAP1E derivatives (pCDNA3.1-AAPs) were transfected as described in Materials and Methods and visualized by their native EGFP fluorescence. AAP1 was immunostained with α-C9 antibody, and nucleoli were immunostained with C23 antibody. Nuclei were stained with DAPI. (E) Relative vector yields using different AAP constructs normalized to wild-type AAP1 (AAP1). (F) Transduction of rAAV1 packaging luciferase produced from different AAP1 constructs as described above. RLUs were normalized to AAP1. Error bars were determined and statistical analyses were performed as described above.