Figure 8

Knockdown of DVL1 or DVL3 reduces proliferation in alveolar rhabdomyosarcoma cells. (A) Heatmap of 22 differentially expressed genes associated with the WNT pathway in fusion-negative (pink) and fusion-positive (various colours) rhabdomyosarcoma biopsies and healthy muscle controls (Normal – orange) (GSE108022). (B) Localisation of DVL1-3 (green) in RH30 ARMS cells exposed to high-serum medium, co-immunolabelled for β-TUBULIN (red) with nuclei counterstained with DAPI (blue). (C) Gene expression profiles for DVL1-3, the proliferation marker CD1 and the early differentiation marker MYOGENIN in RH30 cells exposed to high or low-serum medium. N = 3 independent biological replicates. Significant differences were assessed using a students unpaired two tailed T-test, comparing high-serum to low-serum culture conditions where two asterisks denote p < 0.01. (D) Representative images of proliferating RH30 cells that have incorporated EdU (red) after individual knockdown of each DVL isoform for 48 h, with nuclei counterstained with DAPI (blue). Scale bar represents 100 µm. Quantification of EdU containing cells as a percentage of total nuclei after knockdown of each DVL isoform. Statistically significant differences were calculated using a One-Way ANOVA with Dunett’s post-hoc test, comparing each group with the control (SiCtrl). A significant difference is denoted via two asterisks where p < 0.01. More than 200 nuclei were analysed for each of N = 3 biological replicates.