Fig. 6

MiR-140-3p regulation of BMSC differentiation via Plexin B1. A, B Plexin B1 plays an important role in the interaction between osteoblasts and osteoclasts. The expression of plexin B1 in N-BMSCs and DM-BMSCs was examined using Western blot analysis and qPCR after treatment with miR-140-3p NC/mimics/inhibitor. n = 3 in each group. C Schematic illustration of the sequences for miR-140-3p and the WT or mutated 3′-UTR of plxnb1 mRNA. D Dual Rluc/Fluc luciferase luminescence intensity of plxnb1 WT or mutated 3′-UTR reporter plasmids in HEK293 cells co-transfected with miR-140-3p mimics or miR-NC mimics. E Protein expression levels of p-RhoA, p-ROCK, and p-IRS in N-BMSCs and DM-BMSCs after Sema4D treatment (0, 10, 20, 40 μg/mL). F Levels of p-RhoA, p-ROCK, and p-IRS in N-BMSCs and DM-BMSCs transfected with miR-140-3p NC/mimics/inhibitor after treatment with Sema4D (10 μg/mL) as determined by Western blotting. G Levels of p-RhoA, p-ROCK, and p-IRS in N-BMSCs and DM-BMSCs as measured by Western blotting, after culture with anti-plexin B1 (0, 30, 60, 90 ng/mL) and Sema4D (10 μg/mL). H Levels of Runx-2, ALP, and osterix in N-BMSCs and DM-BMSCs after treatment with Sema4D (0, 10, 20, 40 μg/mL). I qPCR analysis of Runx-2, ALP, and osterix expression levels in N-BMSCs and DM-BMSCs transfected with miR-140-3p NC/mimics/inhibitor and cultured with Sema4D (10 μg/mL). J mRNAs expression levels of Runx-2, ALP, and osterix in N-BMSCs and DM-BMSCs treated with anti-plexin B1 (0, 30, 60, 90 ng/mL) and Sema4D (10 μg/mL). Data represent means ± SD (n = 3 in each group. Western blotting and qPCR experiments were repeated 3 times). *p < 0.05; **p < 0.01; ***p < 0.001