Chromosomal integration by AAV/HM1.24/Neo in DCs. Cells, treated as indicated and as described in “Materials and methods” and sorted for CD83 expression, were further analyzed for chromosomally integrated AAV/HM1.24/Neo viral genomes. Total cellular DNA (0.1 μg) from infected, CD83+-selected, and uninfected cells served as a template in PCR amplification assays using primers targeting the SV40 early promoter of the vector and the cellular repetitive AluI element. The products underwent Southern blotting and were probed with 32P-Neo DNA. The positive control lane contained 100 ng BamHI-digested AAV/HM1.24/Neo plasmid (6.5 kb and 2 kb). Note that multiple Neo-positive bands resulted from the infected cell population, indicating chromosomal integration by the vector, and that multiple vector-positive cell clones were present in the population.
