FIG 4.

Tegument protein UL35 affected ss NID1 levels via the proteasome. HFFs were infected with HCMV (A) or the indicated Ad vector (C, E, and G) at the indicated MOI. At 4 hpi (A, C, and E) or 24 hpi (G), 0.5 μM MG132 (or DMSO vehicle) was added to inhibit proteasomal activity. Cells were harvested at 48 hpi (A, C, and E) or 72 hpi (G). Cell lysates (25 μL) were assayed via Western blotting for ss NID1. Pan-actin was used as a loading control to normalize the band intensity of NID1 for calculations of V/M or Ad viral protein/Adic ratios as described in Materials and Methods and the text and shown below each blot. (B, D, F, and H) Scatter plots of values of normalized NID1 intensity (V/M or Ad viral protein/Adic) ratios from different experiments like those represented in A, C, E, and G, respectively. Horizontal bars represent the means of repeated experiments.