Fig 2: LNCaP and PC3 cell invasion increased after the addition of HS-5 cell CM. (A) LNCaP and PC3 cell invasion increased after the addition of CM from HS-5 immortalized stromal cells. (B) The efficiency of shRNA-mediated A/B knockdown by Western blot. (C) LNCaP and PC3 cell invasion in the presence of HS-5 CM. Decreased cell invasion was observed in the presence of shRNA-mediated WNT5A knockdown in PC3 (P = .011) and LNCaP (P = .013) cells.

Fig 3: Expression of muscle markers in WNT5AOE and WNT5AKD RMS cell lines. Western blot analyses (top) along with the respective protein quantification (bottom) of the muscle markers MYOD, DES and MYOG of the human ARMS cell line RH30 and ERMS cell lines RD and TE671 stably overexpressing WNT5A or stably expressing a WNT5A shRNA to induce a WNT5A knockdown. Cell lines transduced with the respective empty vectors (pBABE or pGIPZ) served as controls. Detection of ß-Actin served as loading control. For protein quantification from =3 independent experiments, values of CTR were set to 1. Bars are mean ± SD; statistical analysis was performed by one-way ANOVA. *indicates significance vs. CTR; #indicates significance between OE and KD. */#P<0.05, **/##P<0.01, ***/###P<0.001, ####P<0.0001. RMS, rhabdomyosarcoma; ARMS, alveolar RMS; ERMS, embryonal RMS; CTR, control; OE, overexpressing; KD, knockdown.

Fig 4: Wnt5a/ROR2 expression is increased in lysates from platinum resistant tumors. A. Western blot analysis of lysates from platinum resistant vs. platinum sensitive patient samples with the indicated primary antibodies. B. Quantification of protein expression for Wnt5a, ROR2, STAT3 and NF-kB based on ß-tubulin levels. Horizontal lines represent the average and errors bars are SEM (standard error of mean). After normalization, samples were quantified based on the highest expression level of all samples.

Fig 5: Wnt5a regulates the expression of ABCG2 gene through FZD7. Data sets of ‘Tumor Pancreatic adenocarcinoma-TCGA-178-rsem-tcgars’, ‘Tumor Pancreatic ductal adenocarcinoma-Yeh-132-custom-4hm44k’ and ‘Tumor Pancreatic ductal adenocarcinoma (ICGC)-Perez-91-complex-ilmnht12v4’ in the R2 database demonstrated the correlation between Wnt5a and FZD7 in pancreatic carcinoma (A-a) correlation between FZD7 and ABCG2 in pancreatic carcinoma (A-b). (B-a) Protein levels of FZD7 in four pancreatic cancer cell lines. (B-b) Protein levels of FZD7 after transient transfection of different siRNA molecules targeting FZD7. (C) rhWnt5a does not upregulate ABCG2 and ß-catenin expression following transfection with siFZD7. (D) ABCG2 protein expression following FZD7 overexpression in Capan-2 cells treated with rhWnt5a. *P<0.05, **P<0.01, ***P<0.001. ABCG2, ATP binding cassette subfamily G member 2; rhWnt5a, recombinant human Wnt5a; FDZ7, Frizzled class receptor 7; siRNA, small interfering RNA; con, control.