Fig 3.

Infectivity of pRCAS-AP and derivatives. (A) DF1 cells were transfected with pRCAS-WT or mutant derivatives (Transfection), and the released virus was used subsequently to infect new cells (Infection) as detailed in Materials and Methods. Cell lysates (upper panels) and supernatant virus (lower panels) were analyzed by SDS-PAGE (8% and 15% gels, respectively) and Western blotting using a polyclonal anti-CA (p27) serum. (B) Duplicate dishes from the transfection illustrated in panel A were stained for alkaline phosphate activity. (C) Analysis of the replication of pseudoknot stem 2 (S2) and stem 3 (S3) destabilization mutants. Cells and virus were processed and analyzed as described for panel A.