Figure 3

Rap1A is a new target of miR-203. (A) The putative miR-203 binding sequence was in the 3′-UTR of Rap1A mRNA. (B) A dual luciferase report assay was performed in HEK-293 T cells cotransfected with psi-CHECK2- Rap1A and miR-203. The dual luciferase report assay of HEK-293 T cells cotransfected with RAP1A-3′-UTR-WT or Rap1A-3′-UTR-MUT and empty vector control or pCDH-miR-203. The Renilla luciferase activity was used as control. (C) Western blot analysis of the endogenous expressions of Rap1A upon forced expression of miR-203. (D) The protein expression of Rap1A in PC-3 and DU145 cells transfected with miR-203 inhibitor or NC was analyzed by western blotting. (E) Quantitative real-time PCR analysis showing the mRNA levels of Rap1A. (F) The relative expression of Rap1A in PCa tissues and matched adjacent normal tissues was determined by qRT-PCR. (G) Plots showing a negative correlation between the relative expression levels of miR-203 and Rap1A in the above samples. * p < 0.05, ** p < 0.01.