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

Fig. 2. MiR-185 silencing promotes primary osteoblast differentiation and mineralization.

Fig. 2

a Cell Counting Kit-8 (CCK-8) assay reflected cell proliferation of osteoblasts derived from wild-type (WT) or miR-185-knockout (KO) mice calvaria. b Alkaline phosphatase (ALP) activity determination in WT or miR-185-KO osteoblasts after cultured with osteoblast induction medium (OIM) for 7 days (n = 3). c Matrix mineralization was quantified in primary osteoblasts after induction in OIM for 14 days (n = 3). d Representative images of ALP staining of WT or miR-185 KO cells after osteoblast induction for 7 or 14 days. Scale bar = 500 μm. e Representative images of Alizarin Red S staining in cells after osteoblast induction for 14 or 21 days. Scale bars = 500 μm. f The primary osteoblasts were cultured in OIM for indicated times. RNA in cells was extracted with TRIzol reagent, and the expression levels of osteoblast marker genes were quantified by real-time PCR (n = 3). g The protein levels of osteoblast marker genes in primary osteoblasts cultured with OIM for 0, 3, and 7 days were analyzed by western blot, and expressed as densitometry normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Data were representative of at least three independent experiments, and were shown as the mean ± S.D (*P < 0.05, **P < 0.01, ***P < 0.001)