FIG. 8.

Rotavirus mRNA half-life increases in infected cells. MA104 cells (5 × 10⁵ cells/assay) were infected (+ virus) or not (− virus) with RRV at an MOI of 3. At 1 hpi, 1 μg of either luciferase (Luc) or gene 10 reporter mRNA was transfected using Lipofectamine 2000 for 2 h. At the indicated times posttransfection, cells were lysed, and the transfected mRNAs were quantified by RT-PCR using primers for luciferase or the reporter gene 10Mut (Table 1), as described in Materials and Methods. The half-lives of the exogenous capped RNAs were calculated by a linear regression analysis of the results from the qRT-PCR at different times posttransfection. The half-life of rotavirus gene 10 mRNA in mock-infected cells was about 12 h, while in infected cells no significant decay was observed during the 12 h of the assay. The luciferase mRNA half-life (240 min) remained the same in uninfected cells and cells infected with rotavirus. Data represent the arithmetic means ± standard deviation of three independent experiments.