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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: DZNep promotes mouse bone defect healing via enhancing both osteogenesis and osteoclastogenesis

Fig. 2

DZNep promoted osteoclastic bone resorption, osteoclast precursor cell fusion, and the expression of osteoclast-specific genes in vitro. A BMMs were seeded onto the hydroxyapatite-coated Osteo Assay plates and cultured with M-CSF (30 ng/mL), RANKL (50 ng/mL), and with 0, 6.25, 12.5, and 25 nM DZNep for 5 days. Scale bar = 200 μm. B Quantification of bone resorption area was performed by ImageJ (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). C The perimeter of the actin ring was measured using ImageJ. D Representative immunofluorescence images for podosome actin belt formation. Scale bar = 20 μm.(*p < 0.05; **p < 0.01; ***p < 0.001). E Expression of the osteoclast-specific genes Trap, Ctsk, and Atp6v0d2 in BMMs treated with 25 nM DZNep, M-CSF (30 ng/mL), and RANKL (50 ng/mL) for 0, 1, 3, and 5 days. F Expression of the osteoclast-specific genes Trap, Ctsk, Atp6v0d2, Calcr, Nfatc1, and c-Fos in BMMs treated with M-CSF (30 ng/mL), RANKL (50 ng/mL), and indicated DZNep concentrations for 5 days. Gene expression was analyzed by real-time PCR. mRNA expression levels were normalized relative to the expression of β-actin (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). Data are expressed as median and interquartile range, n = 3

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