The <i>m154</i> gene ectopically expressed within the MCMVΔm154 genome decreases CD48 surface levels on MCMV-infected macrophages.

<p>(A) Schematic depiction of MCMVm154Ectop, with an expansion of the <i>m153-m155</i> region (represented by an open box) to show the deletion of the <i>m154</i> gene, and of part of the MIE region (represented by a black box) to indicate the location of the <i>m154</i> gene inserted (together with 210 nt of its corresponding putative promoter and 60 nt including its putative polyA signal [pA]) in between the <i>ie2</i> and the <i>m129</i> genes. The major IE promoter (MIEP) is marked. (B) Peritoneal macrophages were mock-infected or infected for 72 h with wt MCMV or MCMVm154Ectop at an moi of 10 and analyzed by flow cytometry for surface expression of CD48 as described in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004000#ppat-1004000-g001" target="_blank">Figure 1</a>. Shaded histograms represent CD48 expression on mock-infected cells, and black line and dotted-line histograms represent CD48 expression on cells from wt MCMV- and MCMVm154Ectop-infected macrophages, respectively. Isotype IgG was used as a negative control (dashed line histograms). A representative experiment out of two performed is shown. (C) Peritoneal macrophages, infected with MCMVm154Ectop at an moi of 10 for 72 h, were fixed, permeabilized, and stained with anti-m154 mAb followed by an anti-mouse IgG Alexa fluor 555, and analyzed as indicated in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004000#ppat-1004000-g004" target="_blank">Figure 4C</a>. Shown are nuclei stained with the DAPI reagent (left panel), cultures stained for m154 (middle panel), and the overlaid image (right panel). Images were captured at 40× magnification.