CD45 surface expression is reduced in MCMV-infected RAW264.7 macrophages.

(A) RAW264.7 cells were either mock infected (open histogram) or infected with MCMVgfp (filled histograms) at an MOI of 3. 24 h p.i. CD45 surface expression was determined by flow cytometry for all cells of the cultures, except dead cells, which were excluded based on 7-AAD staining. Dotted line, isotype control. (B) Localization of CD45 was assessed 24 h p.i. by fluorescence microscopy in uninfected and infected (GFP+) RAW264.7 cells that were fixed, permeabilized and immunostained with a CD45-specific Ab. Cell nuclei were counterstained with Hoechst dye. Scale bars, 10 μm. (C) Schematic representation of the 230-kb MCMV genome (HindIII map), indicating the genes lacking in the respective deletion mutants. (D) RAW264.7 cells were mock-infected (open histograms) or infected (filled histograms) with the indicated deletion mutants, and 24 h p.i. immunostained to analyze CD45 surface levels. Dotted line, isotype control. For (D) gating was on living cells and for samples with infected cells additionally on GFP⁺ cells.