Targeting DNA-PKcs reduced rAAV replication.

<p>293 cells were transfected with DNA-PKcs specific siRNA (DNA-PKcs siRNA) or control siRNA. Two days after transfection, cells were infected with rAAV-UF5 and rHSV, or transfected with pUF5 and pDG. rAAV replication was evaluated by Southern blot analysis as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0015073#pone-0015073-g001" target="_blank">Figure 1</a>. (A) Effect of DNA-PKcs siRNA on rAAV replication by Southern blot after vector infection. Concentration of DNA-PKcs siRNA and control siRNA was 100 pmole. (B) Densitometry analysis of data from Figure A. **, <i>P<0.001</i> for monomer; *, <i>P<0.05</i> for dimer and concatamers when compared with control siRNA group. (C) Dose dependent effect of DNA-PKcs siRNA on rAAV DNA replication. (D) Effect of DNA-PKcs siRNA on rAAV after vector and helper plasmid transfection. Left panel, without DpnI digestion; Right panel after. Note: DpnI digestion removes transfected plasmid DNA and shows all <i>de novo</i> replicated rAAV forms (monomer, dimer and concatamers).