FIG. 3.

AAV RNA accumulation following helper virus preinfection. (A) P5/19 products accumulate prior to other AAV RNAs in 293 cells preinfected with Ad. The RP probe was used to protect 10 μg of total RNA exactly as for Fig. 2 except that 293 cells were preinfected with Ad (MOI = 2 to 5) for 6 or 12 h (indicated at the bottom). Total RNA was isolated at the times indicated after AAV infection at time zero. u, unspliced; s, spliced. (B) P5 products accumulate prior to P19 products in 293 cells preinfected with Ad. The SB probe was used to protect the same RNA samples used for experiments shown in panel A. Asterisks indicate likely breakdown products of P5 protected fragments and were excluded from quantification. (C) There is a shift to the relative predominance of spliced AAV RNA species in 293 cells preinfected with Ad. The DH probe was used to protect the same RNA samples as used for experiments shown in panels A and B. Unspliced RNAs and spliced RNAs utilizing either A1 or A2 are indicated.