Fig. 5: Elevation of ROR2 suppressed PI3K–AKT signaling via activation of PIAS3.

(a) Proteins expression level of ROR2, PIAS3, phospho-AKT S473, phospho-AKT T308, and total AKT in PC-3 cells with or without ROR2 overexpression and PIAS3 siRNA knockdown in the presence or absence of 50 ng/ml EGF for 24 h treatment was determined by the western blotting assay. Expression of β-actin was used as loading control. (b) Cell migration of PC-3 cells with or without ROR2 overexpression and PIAS3 siRNA knockdown in the presence or absence of 50 ng/ml EGF for 24 h treatment was determined by the wound healing assay. Expression level of hsa-miR-199a-5p in control PC-3 cells or PC-3 overexpressing ROR2 cells (c) or in control DU-145 cells or DU-145 overexpressing ROR2 cells (d) was determined by qRT-PCR. Expression of U6 snRNA was used as loading control. PC-3 (e) or DU-145 (f) cells transfected with control vector or pCMV-ROR2 vector with or without overexpression of miR-199a-5p were cultured for 72 h. Migration ability of these PCa cells was determined by transwell assay. Asterisks *, **, and *** represent statistically significant difference p < 0.05, p < 0.01, and p < 0.001, respectively, between the two groups being compared. (g) Expression level ROR2 and PIAS3 proteins in PC-3 cells transfected with control vector or pCMV-ROR2 vector with or without overexpression of miR-199a-5p was determined by the western blotting assay. The β-actin was used as loading control.