FIG. 6.

Effect of pZ cotransfection on RNA replication. Subconfluent cultures of CV1 cells previously infected with vTF7-3 were transfected with minigenome (A), miniantigenome (B), or pS-GenCAT (C) together with pN with or without pL. When pZ was included, its amount is indicated in nanograms. Total RNA (A, lanes 5 to 7) was obtained at 24 h posttransfection, whereas nucleocapsid-associated RNA (Nc RNA; A and B, lanes 1 to 4; and C, lanes 1 to 5) was immunoselected at 40 h posttransfection. Purified RNAs were then analyzed by Northern blotting. Duplicate blots were hybridized either with an antisense CAT riboprobe [CAT(−)] (upper panels) or a sense CAT riboprobe [CAT(+)] (lower panels). Arrows show the positions of the RNAs synthesized by the TV polymerase or the T7 polymerase (T7 transcript). “kb” indicates the sizes of RNA markers resolved in parallel. In panels A and B, films were exposed for 20 h, with the exception of lanes 5 to 7 (A), which were exposed for 2 h. In panel C, films were exposed for 4 days.