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Osteoblast differentiation is tightly regulated by several factors including microRNAs (miRNAs). In this paper, we report that pre‐mir‐15b is highly expressed in differentiated osteoblasts. The functional role of miR‐15b in osteoblast differentiation was determined using miR‐15b mimic/inhibitor and the expression of osteoblast differentiation marker genes such as alkaline phosphatase (ALP), type I collagen genes was decreased by miR‐15b inhibitor. Runx2, a bone specific transcription factor is generally required for expression of osteoblast differentiation marker genes and in response to miR‐15b inhibitor treatment, Runx2 mRNA expression was not changed; whereas its protein expression was decreased. Even though Smurf1 (SMAD specific E3 ubiquitin protein ligase 1), HDAC4 (histone deacetylase 4), Smad7, and Crim1 were found to be few of miR‐15b's putative target genes, there was increased expression …