Figure 2.

P5-associated AAV contaminants are transcriptionally active after infection

(A) Sequential RNA-DNA nuclear FISH assay of mouse hepatocytes 1 week after 8.4 × 10¹¹vg AAV8ssHLPhFVIII injection (top) or uninfected C57/BL6J (bottom). Red indicates probe detecting contaminant Rep-Cap plasmid sequences. Blue indicates DAPI staining. White arrows identify contaminant sequence hybridization. RNA FISH (left), RNA FISH after RNase treatment (middle), DNA FISH (right). (B) Pie chart of NGS read contributions mapping to produce plasmid contaminant 4 months after 8.4 × 10¹¹vg injection with AAV8ssHLPhFVIII from mouse liver DNA (top) and mouse liver RNA (bottom). Reads mapped as in (Figure 1D).