Figure 4

Constitutive overexpression of DVL1 or DVL3 enhances myoblast proliferation. (A) Gene expression levels of DVL1 (left), DVL2 (middle) and DVL3 (right) in proliferating human C25 DVL1⁺, DVL2⁺ and DVL3⁺ myoblasts with stable lentiviral-mediated overexpression of each isoform. N = 4 biological replicates. Significant differences were calculated using a One-Way ANOVA with Dunett’s post-hoc test, comparing each group with the control (pUltra) where an asterisk denotes p < 0.05, two asterisks denote p < 0.01 and three asterisks is p < 0.001. (B) Representative images of proliferating C25 DVL1⁺, DVL2⁺ or DVL3⁺ myoblasts overexpressing the respective DVL isoform, immunolabeled for GFP from the lentiviral backbone (indicative of DVL levels) (green), with a DAPI nuclear counterstain (blue). Myoblasts with incorporated EdU (red) are also shown, after a 2 h EdU pulse performed 24 h after seeding. Scale bar represents 100 µm. (C) Quantification of C25 or 16U DVL1⁺, DVL2⁺ or DVL3⁺ myoblasts containing EdU as a percentage of total nuclei. More than 200 nuclei were analysed for each N = 4 biological replicate. Data is represented as mean ± SD with significant differences calculated using a One-Way ANOVA with Dunett’s post-hoc test, comparing each group with the control (pUltra), where one asterisk indicates p < 0.05, two asterisks indicate p < 0.01 and 3 asterisks p < 0.001.