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. 2002 Dec;76(24):12435–12447. doi: 10.1128/JVI.76.24.12435-12447.2002

FIG. 2.

FIG. 2.

Northern blot analysis of AAV5 RNA. (A) RNA was isolated at 36 to 40 h postinfection from 293 cells infected (I) with AAV5 (15 IU/cell) and Ad5 (5 PFU/cell) (lane 1) or with AAV2 (10 IU/cell) plus Ad5 (5 PFU/cell) (lanes 3 and 5) or from uninfected (U) 293 cells (lanes 2, 4, and 6). Northern blots were hybridized to either an AAV5 whole-genome probe (lanes 1 and 2), an AAV2 whole-genome probe (lanes 3 and 4), or an AAV2 rep gene probe spanning AAV2 nt 810 to 965, which overlap the P19 promoter (lanes 5 and 6), as indicated. The sizes of the RNA species are shown to the left of each panel. The identities of the AAV2 transcripts are as follows: unspliced P5-generated RNA (4.3 kb), unspliced P19-generated RNA (3.6 kb), spliced P19-generated RNA (3.3 kb), unspliced P40-generated RNA (2.6 kb), and spliced P40-generated RNA (2.3 kb). (B) RNA was isolated at 36 to 40 h postinfection from 293 cells infected (I) with AAV5 (15 IU/cell) plus Ad5 (5 PFU/cell) (lane 1). Northern blots were hybridized to either a whole AAV5 genomic probe (lane 1), a subfragment of the AAV5 rep gene spanning AAV5 nt 1434 to 1654 (lane 2), or a subfragment of the AAV5 cap gene spanning AAV5 nt 3359 to 3599 (lane 3), as indicated. The sizes of the bands are shown on the left. AAV5 rep and cap gene probes contain similar number of labeled adenosines.