Figure 7

WNT5A affects angiogenic processes in malignant melanoma cells. (A) Quantification of total length of tubes (left), quantification of number of tubes (right), n = 4. Error bars represent SEM. * = p <0.05, ** = p <0.01 (B) MS1 cells grown together with HTB63 treated with DMSO or Bapta-AM, (left), total length of tubes, (right), number of tubes, n = 3. Error bars represent SEM. * = p <0.05 ** = p <0.01 by paired Student’s t-test. (C) MS1 cells that had been stimulated with exosomes isolated from carrier- or rWNT5A-treated Mewo cells, or with exosome-depleted supernatant from the corresponding samples, to analyze the relative total tube length induction, number of tubes, n = 6. Error bars represent SEM. ** = p <0.01 by paired Student’s t-test. (D) Q-PCR of mRNA from MS1 cells that had been stimulated with exosomes isolated from carrier- or rWNT5A-treated Mewo cells to analyze angiogenesis related genes using mouse specific primers. Both ALK1 (black bars) and Endoglin (white bars) expression was enhanced by rWNT5A induced exosomes (exosome fraction; left) whereas the exosome depleted supernatant fraction showed a loss of function as compared to total supernatant (exosome depleted fraction; right) (E) Correlation between WNT5A and the angiogenesis marker endothelial cell-selective adhesion molecule (ESAM) mRNA expression levels in a set of 223 primary malignant melanomas [24]. R = 0.31; Spearman’s rank test P = 7.7 × 10^-7.