Figure 3

PTCH1 was a direct target of miR-212 in PDAC cells. (A) The miR-212 biding site predicted in the 3′UTR of PTCH1 mRNA. Mutant was generated at the seed region of PTCH1 3′UTR as indicated by the underline. A 3′UTR fragment of PTCH1 mRNA containing wild-type (WT) or mutant (Mut) of the miR-212 binding sequence was cloned into the downstream of the luciferase gene in pGL3b vector. (B) PANC-1 cells were transfected with pGL3b reporter vectors containing either wild-type (WT) or mutant (Mut) PTCH1 3′UTR with either miR-212 mimic or miR-212 inhibitor, or negative control (NC). Luciferase activity was determined 48 h after transfection. (C) PTCH1 mRNA was detected by qRT-PCR in PANC-1 cells transfected with miR-212 mimic or NC or miR-212 inhibitor. (D) The protein levels of PTHC1 were examined by western blot assays transfected with miR-212 mimic or miR-212 inhibitor or NC. *P < 0.05.