Research Paper Volume 11, Issue 21 pp 9442—9460

Long noncoding RNA DNM3OS promotes prostate stromal cells transformation via the miR-29a/29b/COL3A1 and miR-361/TGFβ1 axes

Figure 4. miR-29a and miR-29b directly bind the DNM3OS and COL3A1 3'UTR to negatively regulate their expression (A) PrSCs were transfected with si-DNM3OS and examined for the expression of miR-29a/29b by real-time PCR. (B) miR-29a/29b overexpression or inhibition conducted in PrSCs by transfection of miR-29a/29b mimics or inhibitor and confirmed by real-time PCR. (C) PrSCs were transfected with miR-29a/29b mimics or inhibitor and examined for the expression of DNM3OS by real-time PCR. (D) PrSCs were transfected with miR-29a/29b mimics or inhibitor and examined for the protein levels of COL3A1. (E) A schematic diagram showing the predicted binding sites between miR-29a/29b and DNM3OS or COL3A1. Wild- and mutant-type DNM3OS or COL3A1 3'UTR luciferase reporter vectors were constructed. Mutant-type vectors contained a 7-bp mutation in the predicted miR-29a/29b binding site. (FG) 293T cells were cotransfected with these vectors and miR-29a/29b mimics or inhibitor and examined for luciferase activity. (H) PrSCs were cotransfected with si-DNM3OS and miR-29a/29b inhibitor and examined for the protein content and distribution of COL3A1 by IF staining (scale bar: 50 μM). **P<0.01.