Figure 6

Mutation of the NLS sequence in DVL1 or DVL3 leads to reduced proliferation. (A) Schematic of the three conserved domains (DIX, PDZ, DEP) in DVL proteins, together with the positions of the nuclear localisation sequence (NLS) and the nuclear export sequence (NES), together with the changes to key amino acid residues to destroy the NLS (red text). (B) Representative images of proliferating C25 DVL1⁺, DVL1-mNLS⁺, DVL3⁺, DVL3-mNLS⁺ and control (Ctrl) myoblasts, with incorporated EdU (red), immunolabelled for GFP (green) from the lentivirus (correlated with DVL expression) and DAPI nuclei counterstain (blue). EdU pulse was performed 24 h after seeding stably overexpressing C25 myoblast lines. Scale bar represents 100 µm. Quantification was performed for 4 biological replicates, and significant difference was calculated using a One-way ANOVA with Dunnett’s post hoc test, comparing each group with the control, where an asterisk denotes p < 0.05 and 3 asterisks denotes p < 0.001. (C) Representative images of DVL3⁺ (left) or DVL3-mNLS⁺ (right) C25 myoblasts. Red arrows point to an unusually round, and an uncharacteristically elongated, DVL3-mNLS⁺ myoblast.