FIG. 6.

Inhibition of MHV gene expression by UO126. (A) Inhibition of viral RNA synthesis by UO126. DBT cells were infected with MHV-JHM at an MOI of 2 and treated with UO126 or DMSO in the presence of actinomycin D (10 μg/ml). At 2 h p.i., [³H]uridine was added to the medium at a concentration of 50 μCi/ml. The viral RNAs were isolated at various time points p.i. as indicated and were precipitated by TCA. The radioactivity levels of the TCA precipitates were determined by a scintillation counter and are expressed as counts per minute. The data indicate the means of three separate experiments. Error bars denote the standard deviations. (B) Inhibition of viral protein synthesis by UO126. DBT cells were infected with MHV-JHM at an MOI of 2, treated with UO126 [UO126 (+)] or DMSO [UO126(−)] at 1 h p.i., and labeled with [³⁵S]methionine (50 μCi/ml) at various time points for 30 min. Cell extracts were prepared at the end of the 30 min labeling. One-eighth of each sample with equivalent cell numbers was analyzed by SDS-10% PAGE, and the gel was autoradiographed. Arrows indicate the position of three predominant MHV structural proteins (S, N, and M).