Human Cytomegalovirus Infection Dysregulates the Canonical Wnt/β-catenin Signaling Pathway
Figure 3
HCMV induces degradation of β-catenin.
(A) Lysates were collected from HFFs infected with HCMV-TR (MOI of 1–2) at the indicated times post infection and analyzed for β-catenin expression by Western blot. β-actin served as a loading control. β-catenin protein levels were quantitated by densitometric analysis using ImageJ software (n = 3) and normalized to mock-infected cells at each time point, which was set to a value of 1.0. Data are presented as the mean ± SEM of 3 independent experiments. **p<0.01 (B) Membrane, cytoplasmic and nuclear protein-enriched cell fractions were prepared from HFFs 48 hr after infection with HCMV-TR (MOI of 1–2), and analyzed for β-catenin expression by Western blot. Caveolin-1, GAPDH, and histone H4 served as loading controls for the membrane, cytoplasmic and nuclear fractions, respectively. β-catenin protein levels were quantitated by densitometric analysis using ImageJ software. (C) β-catenin mRNA expression in HCMV-TR-infected (MOI of 1–2) or mock-infected HFFs at 24, 48 and 72 hr post infection were analyzed by qRT-PCR. Relative β-catenin mRNA levels in virus-infected samples at each timepoint were normalized to GAPDH mRNA levels and expressed as fold change relative to the corresponding uninfected control. Data are presented as mean +/− SEM of 3 independent experiments. *p<0.05.