Fig. 2

Prostate cancer cells-derived PKD2/3 promote chemotactic migration of mast cells and endothelial cells tube formation. a Conditional medium (CM) from PC-3 M or DU145 cells transiently transfected with si-CTL, si-PKD2, si-PKD3 was used to be chemoattractant for P815 mast cell migration by transwell assays. ***p < 0.001, **p < 0.01 versus si-CTL by one-way ANOVA. b Silencing of PKD2 and PKD3 verification by Western blotting (left panel). c VEGF mRNA level was assessed by qPCR in P815 mast cells induced by conditional medium from DU145 cells transfected with siRNA of PKD2 or /and PKD3 (right panel). *p < 0.05, **p < 0.01 versus si-CTL by one-way ANOVA. d Knockdown efficiency were verified by Western blot in PC-3 M(up) or P815 mast cells(bottom). Cells were transiently transfected with siRNA of indicated genes using lipofectmine3000. e HUVEC cells were treated with conditional medium from co-cultured of P815 mast cells transfected with si-CTL or si-VEGF and PC-3 M cells transfected with si-CTL, si-PKD2, si-PKD3 or si-PKD2 + 3. Tube formation of HUVEC cells were visualized by phase contrast inverted microscope (100×). f Tube formation was assessed the number of nodes per image was quantified and analyzed by two way ANOVA, **p < 0.01