FIG 3.
The UL133/8 locus is required for high-level accumulation of mRNA and protein for IE2 and for two viral proteins involved in DNA synthesis and for a UL97 inhibitor to influence their accumulation. (A to C) mRNA abundance at 48 hpi for UL122 (IE2), UL44, and UL57 is shown for infections of fibroblasts with either WT or UL133/8NULL at an MOI of 1 in the presence of 2 μM MBV or 0.1% DMSO. mRNA abundance is shown relative to that in DMSO control-treated WT infections, which were set to 1.0. The results represent the averages from two independent biological replicates; a one-way analysis of variance with Tukey's posttest was used to evaluate statistical significance. (D) Fibroblasts were infected with WT or UL133/8NULL at an MOI of 1 or mock infected (lanes m) and maintained in the presence or absence of MBV. A series of protein lysates was collected at the indicated time points, and expression of the specified viral proteins was monitored by Western blotting. (E) Fibroblasts were infected with WT or TB40/E Δ97 (Δ97) at an MOI of 1 or mock infected, and lysates were collected at 6, 16, 24, and 48 hpi. (F and G) Bands from detection of IE2, UL44, and UL57 at 48 hpi were quantified and normalized to the beta-actin signal. Results are shown relative to the signal intensity from DMSO-treated WT-infected samples, which was set to 100. (F) Results from three independent biological replicates of the conditions compared in panel D. (G) Results from two independent replicates of the experiment whose results are shown in panel E. A one-way analysis of variance with Tukey's posttest was used to determine statistical significance in panel F, and the Student t test was used for panel G. Error bars indicate SEMs. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.