Fig. 7.

MiR-144-5p impairs BMSCs osteogenesis differentiation via targeting Smad1. a Venn diagram of the targeting genes of miR-144-5p predicted by microT, TargetScan, miRDB, and miRmap. b–d Western blotting and qRT-PCR analysis of Smad1 protein and mRNA levels, respectively, in BMSCs transfected with mimic-NC, miR-144-5p-mimic, inhibitor-NC or miR-144-5p-inhibitor e WT and mutated miR-144-5p recognition sites within the Smad1 3’ UTR were constructed for the dual-luciferase reporter assay and the results demonstrated Smad1 as a direct target gene for miR-144-5p. f–h After BMSCs were con-transfected with miR-144-5p-inhibitor or si-Smad1; the mRNAs and proteins of Smad1, RUNX2, ALP, collagen Ι and OCN were determined using qRT-PCR and Western blotting. I the mineralization in BMSCs following different treatments was measured via Alizarin red staining and ALP staining. j–k The statistical data of Alizarin red staining and ALP staining. Data are presented as the mean±SD, and all experiments were repeated three times. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.001