Research Paper Volume 15, Issue 14 pp 7124—7145

Figure 5. PRLR is a downstream target of miR-181a-5p in mouse mandibular BM-MSCs. (A) Normal mandibular BM-MSCs were transfected with NC-mimic or miR-181a-5p. At 48 hours after transfection, the mRNA levels of potential miR-181a-5p targeted candidate genes were quantitated by qPCR. (B) Schematic diagram shows the matching base pairs between miR-181a-5p and the wild type (WT) or mutated (MUT) 3′UTR of PRLR mRNA. The luciferase activity of the reporter vectors was detected in mandibular BM-MSCs at 48 hours after co-transfection of the plasmid expressing wild type or mutant 3′UTR of PRLR mRNA together with the NC mimic or the miR-181a-5p mimic. (C, D) Functional inhibition (C) or overexpression (D) of miR-181a-5p enhanced or reduced PRLR protein expression in mandibular BM-MSCs, respectively. The representative western blot images are shown, and the relative band intensity of PRLR by densitometry was summarized. (E, F) Mandibular-BM-MSCs (but not blood cells) from mice with periodontitis displayed a lower expression level of PRLR mRNA (E) or protein (F) than that from normal controls. The mRNA and protein levels were quantitated by qPCR and western blot assays, respectively. n = 3 for each group; ^*P < 0.05, ^***P < 0.001, between the indicated groups.