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Fig. 7 | Stem Cell Research & Therapy

Fig. 7

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

Fig. 7

DZNep promoted osteoblasts formation through the EZH2-H3K27me3-Wnt4 axis. A EZH2 expression levels in BMSC treated with 0, 25, 50, 100 nM of DZNep combined with osteoblast-cultured medium for 24 h. B The gray levels of EZH2 were quantified and normalized to β-actin using ImageJ. C H3K27me3 expression levels in BMSC treated with 0, 25, 50, 100 nM of DZNep combined with osteoblast-cultured medium for 24 h. D The gray levels of H3K27me3 were quantified and normalized to β-actin using ImageJ. E Expression of the wnt signaling pathway-specific genes Wnt4 in BMSC treated with 0, 25, 50, 100 nM of DZNep combined with osteoblast-cultured medium for 24 h. F Expression of the wnt signaling pathway-specific genes Wnt4 in BMSC treated with or without 100 nM of DZNep combined with osteoblast-cultured medium for 7 days. G Expression of the osteoblast-genes Runx2, Ocn, Coll-1a, and Opn in BMSC treated with or without 100 nM of DZNep combined with osteoblast culture medium for 0 and 7 days. H Expression of the osteoblast-genes Runx2, Ocn, Coll-1a, and Opn in BMSC treated with 0, 25, 50, 100 nM of DZNep combined with osteoblast-cultured medium for 24 h. I BMSCs cells were treated with negative control or different shRna of EZH2, and the levels of EZH2 were determined. J BMSCs were treated with negative control or shRna-2 of EZH2 and stimulated by osteoblast-cultured medium for 7 days. Then the ALP staining was performed after cells were fixed with 4% paraformaldehyde. Scale bar = 20 μm. K Percentage of ALP+ area of total area was quantified by ImageJ. L BMSCs cells were treated with negative control or different siRna of Wnt4, and the levels of Wnt4 were determined. M BMSCs were treated with siRna-NC, siRna-NC with DZNep or siRna-3 of Wnt4 with DZNep and stimulated by osteoblast-cultured medium for 7 days. Then the ALP staining was performed after cells were fixed with 4% paraformaldehyde. Scale bar = 20 μm. N Percentage of ALP+ area of total area was quantified by ImageJ. O Expression of the osteoblast-specific genes Alpl, Ocn, Coll-1a, Opn in BMSCs treated with siRna-NC, siRna-NC with DZNep or siRna-3 of Wnt4 with DZNep and stimulated by osteoblast-cultured medium for 7 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 = 3–5

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