FIG 2.

HCMV infection upregulates A3G in HFFs. (A) Lysates were prepared at the indicated time points and subjected to Western blot analysis for A3G, IEA, and α-tubulin (left panel). A3G protein was subjected to densitometry and normalized to α-tubulin (*, P < 0.05; ***, P < 0.001; one-way ANOVA followed by Bonferroni's posttests, for comparison of infected versus mock-infected cells) (right panel). (B) FRET assay to measure A3G deaminase activity. The average and standard deviation were calculated from three independent experiments (**, P < 0.01; ***, P < 0.001; one-way ANOVA followed by Bonferroni's posttests, for comparison of infected versus mock-infected cells). RFU, relative fluorescence units. (C) HFFs were infected with HCMV at an MOI of 1 or left uninfected (mock) and subjected to immunofluorescence analysis at the indicated time points. A3G (green)/IEA (red) were visualized using primary antibodies followed by secondary antibody staining in the presence of 10% HCMV-negative human serum. Nuclei were counterstained with DAPI (blue). Images were acquired at ×63 magnification, and representative pictures are shown.