FIG. 6.

The NS1 protein does not affect encapsidation of plasmid-supplied or RSV-amplified C2 minigenome. 293 cells were infected with vTF7-3 and transfected with 50 ng of C2 minigenome, 0 (−NS1) or 50 ng (+NS1) of NS1, and the indicated support plasmids in the same amounts as in Fig. 3. The M2 plasmid was that containing both ORFs. Lysates were prepared 40 h postinfection and were either mock treated (lanes 1, 3, 5 and 7, both panels) or treated (lanes 2, 4, 6, and 8, both panels) with micrococcal nuclease (MCN), after which the remaining RNA was isolated. The RNAs were analyzed by Northern blot hybridization with a positive-sense RSV-CAT riboprobe, which would detect minigenome that is plasmid encoded as well as that which is generated by RSV-mediated replication. Some of the plasmid-encoded minigenome accumulates in a form (labeled minigenome w/ ribozyme) that is slightly larger than the minigenome because it contains uncleaved ribozyme at its 3′ end.