Fig. 3: OH2 reduces β-catenin levels and transcriptional activity.

a H1299 and A549 cells were infected with or without OH2. Cells were lysed and the samples were subjected to western blot analysis (left). The quantification data shown are means ± SEM from three independent experiments (right). Statistical analyses were performed using one-way ANOVA with Tukey’s post hoc test. *p < 0.05; **p < 0.01. b H1299 and A549 cells were infected with or without OH2. And the samples were subjected to qRT-PCR analysis. Data shown are means ± SEM from three independent experiments. Statistical analyses were performed using one-way ANOVA with Tukey’s post hoc test. **p < 0.01, ***p < 0.001, ****p < 0.0001. c MG132 does not affect the β-catenin inhibition of OH2. H1299 were treated with OH2 in the absence or presence of MG132 (10 μM). Western blot analysis (left) and quantification (right) of β-catenin were performed. The quantification data shown are means ± SEM from three independent experiments (right). d OH2 reduces β-catenin protein levels in both cytoplasmic and nuclear fractions: H1299 cells were infected with OH2 or mock infection, followed by treatment with Wnt3a for 6 h, followed by nuclear fractionation and immunoblotting. The band density was quantified using ImageJ software. Data are represented as means ± SEM from three independent experiments. e The effects of OH2 on the expression and distribution of β-catenin were explored by IF labeling in the indicated cells with or without Wnt3a (100 ng/mL) stimulation. Magnification ×1000. f OH2 inhibited the transcriptional activity of β-catenin. H1299 and A549 cells infected with OH2 were transfected with TOP-Flash or FOP-Flash. Transfection efficiency was normalized by co-transfection with pRL-TK. Luciferase activity was measured 48 h post transfection by the dual-luciferase assay. Data are shown as the mean ± SEM. g OH2 negatively regulates the expression of MMPs by qRT-PCR analysis. Data shown are mean ± SEM of three independent experiments. Statistical analyses were performed using one-way ANOVA with Tukey’s post hoc test. **p < 0.01; ***p < 0.001; ****p < 0.0001. h Overexpression of UL41 blocks the transcriptional activity of wild type β-catenin: H1299 cells were transfeted with OH2 viral proteins, followed by dual-luciferase assay. Data are represented as means ± SEM from three independent experiments. Statistical analyses were performed using Student’s t test in (c, d, f, h). ns not significant; *p < 0.05; ***p < 0.001; ****p < 0.0001.