FIG. 1.

Kinetics of transcription and replication of the viral RNA. Confluent MA104 cells were infected with RRV at an MOI of 0.1 by adsorbtion for 30 min at 4°C and then for 30 min at 37°C; the noninternalized virions were washed off with 3 mM EGTA in PBS, and MEM at 37°C was added. At the indicated times postinfection, RNAs were Trizol extracted and quantified by RT-PCR as described in Materials and Methods, or with 0.1% Triton X-100 for infectious virus titration. (A) RT-PCR of the negative strand of genes 1, 6, and 10. (B) Quantitation of the mRNAs corresponding to viral segments 1, 6, and 10. (C) RT-PCR of the negative strand of RNA segment 10 and titration of infectious particles by an immunoperoxidase focus assay as described in Materials and Methods. In panels A and B the qRT-PCR results are expressed as the increase relative to the amount of mRNA or negative strand from RNA segment 10 accumulated at 12 hpi, which was taken as 100%. Data shown represent the arithmetic means ± standard deviation of three independent experiments. In panel C data are expressed as percentage of total RNA synthesis or infectious particles produced at 12 hpi, which was taken as 100%. Data shown represent the arithmetic means ± standard deviation of three independent experiments.