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Fig. 4 DIPQUO promotes β-catenin accumulation and activity. C2C12 cells were treated with DIPQUO for (A) time periods and (B) doses indicated in time-course and dose–response analyses to examine β-catenin expression levels by Western blotting in whole-cell extracts. Associated graphs below show expression levels compared with DMSO-treated controls. Relative β-catenin levels are reported as the means ± SD; ∗p < 0.05 and ∗∗p < 0.01 in unpaired two-tailed Student's t test. Ordinary one-way ANOVA yielded F = 3.237 and p = 0.0225 in panel A and F =3.170 and p = 0.0168 in panel B. C, luciferase activity assay using transient expression of GFP-T-cell factor/lymphoid enhancer-binding factor lentivirus (MOI = 20) in 293T cells, followed by treatment with DMSO, 10 μM CHIR or 10 μM DIPQUO. The scale bar represents 50 μm. Luciferase activity normalized to DMSO-treated samples is reported as the mean ± SD; ∗∗∗p < 0.001 in unpaired two-tailed Student's t test. Ordinary one-way ANOVA yielded F = 67.84 and p < 0.0001. Representative images from experiments using at least three biological replicates are shown for all figure panels. MOI, multiplicity of infection; CHIR, Wnt signaling activator CHIR99021.