Figure 5.

COL1A1 is a direct target of miR-625-5p. (A) Schematic representation of the 3’-UTRs of COL1A1 containing two putative miR-625-5p binding sites. These two binding sites are indicated as 1 and 2 and are shown with red lines. The seed locations of miR-625-5p are indicated with red font, while the mutated locations are indicated with blue font. (B-D) COL1A1 can be targeted by miR-625-5p at site 1. The coding sequences of COL1A1 and its wild-type (WT) and mutated (Mut1 and 2) 3’-UTRs were cloned into the pCDNA3 vector. Then, the following combinations of plasmids were transfected into hNPC cells: miR-NC; miR-NC + COL1A1-3’-UTR^WT; miR-NC + COL1A1-3’-UTR^Mut-1; miR-NC + COL1A1-3’-UTR^Mut-2; miR-625-5p-mimic; miR-625-5p-mimic + COL1A1-3’-UTR^WT; miR-625-5p-mimic + COL1A1-3’-UTR^Mut-1; anti-miR-625-5p; anti-miR-625-5p + COL1A1-3’-UTR^WT; anti-miR-625-5p + COL1A1-3’-UTR^Mut-1; and anti-miR-625-5p + COL1A1-3’-UTR^Mut-2. After transfection for 48 hr, cells were collected to detect (B) miR-625-5p, (C) COL1A1 mRNA and (D) COL1A1 protein levels. *P < 0.05 and ***P < 0.001.