FIG. 4.

PF-429242 does not affect arenavirus RNA replication and gene expression. (A) PF-429242 does not affect levels of arenavirus genome RNA replication. (Ai) Schematic of the experiment. BHK-21 cells were infected with LCMV (MOI of 3). At 1.5 hpi, the virus inoculum was removed, cells were washed with PBS, and fresh medium containing PF-429242 (0, 1, 10, or 30 μM) was added. At 1.5 and 13.5 hpi, total cellular RNA was isolated. (Aii) Levels of S genome RNA. RNA (0.5 μg) from each sample was analyzed by RT-qPCR. Copy numbers of S genome RNA were normalized on the basis of GAPDH mRNA copy numbers. Data are averages and standard deviations from three independent experiments. (B) Effect of PF-429242 on LCMV minigenome expression. BHK-21 cells were transfected with pC-T7, pMG-CAT, pC-NP, and pC-L under previously described conditions (17, 18, 26, 29, 30), and 5 h after transfection, the transfection media were replaced with fresh media containing PF-429242 (0, 1, 10, or 30 μM). At 48 h posttransfection, cell lysates were prepared for CAT ELISA. The CAT activity obtained from vehicle (DMSO)-treated cells was set to 1.0 to normalize CAT activities from the other samples. Data are averages and standard deviations from three independent experiments.