FIG. 5.

All AAV RNAs are transported efficiently to the cytoplasm. (A) Total and cytoplasmic RNAs purified at 16 and 24 h after AAV-Ad coinfection were examined using the RP probe in duplicate to control for experimental error. Each species of AAV RNA (indicated on the right [u, unspliced; s, spliced]) accumulates in the cytoplasm in a ratio similar to that found in total RNA preparations. Each species of AAV RNA is efficiently polyadenylated. The RP (B), SB (C), and DH (D) probes were used to protect 10 μg of total RNA purified from AAV-Ad-coinfected 293 cells and poly(A)⁺-containing RNA purified from 20 μg of the same total RNA preparations, as described in Materials and Methods. The ratio of AAV RNA species (indicated on the right of each panel) in poly(A)⁺-purified RNA is indistinguishable from that of total RNA pools for each probe. The asterisk indicates excess undigested DH probe.