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. 2019 Feb 20;10(3):172. doi: 10.1038/s41419-019-1428-1

Fig. 6. Biglycan (Bgn) is the direct target of microRNA-185 (miR-185).

Fig. 6

a Osteoblasts were cultured with osteoblast induction medium (OIM) for the indicated number of days, and Bgn expression was examined by real-time PCR (n = 3). b Luciferase activity was detected 48 h after transfection. c, d MiR-185 mimic and negative control (NC), miR-185 inhibitor and inhibitor NC were transfected into MC3T3-E1 cells, and the messenger RNA (mRNA) expressions of miR-185 and Bgn were examined using real-time PCR (n = 3). e Forty-eight hours after miRNA transfection, the protein level of Bgn in MC3T3-E1 cells was detected by western blot. f Construction of psiCHECK2-Bgn-3′-UTR-WT or mutant plasmids. Schematic of the potential target sites in mouse Bgn 3′-UTR were shown, and two kinds of mutations were generated in the miR-185-5p complementary sites in the reporter vector as indicated. g Normalized luciferase activity of the two mutant reporter vectors after miR-185 overexpression. Data were shown as mean ± SD (*P < 0.05, **P < 0.01, ***P < 0.001). UTR, untranslated region, WT wild type.