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. 2018 Jun 15;8:9225. doi: 10.1038/s41598-018-27514-x

Figure 2.

Figure 2

MiR-10a-3p overexpression promotes ossification of posterior ligament cells. The osteogenic properties of PLL and OPLL ligament cells were analyzed using Alizarin Red staining (A) or alkaline phosphatase staining (B) at different time points of osteo-induction. The colorimetric quantification were shown in the right panels. *P < 0.05, **P < 0.01, n = 3, t-test. The effect of miRNAs overexpression on osteogenic properties of PLL ligament cells were analyzed using Alizarin Red staining (C) or alkaline phosphatase staining (D) at 12 days after osteo-induction. The colorimetric quantification were shown in the right panels. *P < 0.05, **P < 0.01, n = 3, t-test. (E) Real-time PCR analysis of ossification related genes in each treatment group (n = 3) of induced PLL ligament cells. Blank represents osteo-induced cells with no treatment, while NC represents overexpression of a scramble Agomir served as negative control. *P < 0.05, **P < 0.01, t-test. (F) Western blot analysis of ossification related genes in each treatment group (n = 2), uncropped images were shown in the Supplementary Fig. 5. The error bars represents ± S.D.