Fig. 6

The effects of DZNep on osteoblastogenesis and co-cultivation system. A Cell viability of DZNep-treated BMSC was tested by CCK-8 at 12, 48, and 96 h. B BMSC were treated with osteoblast-cultured medium and with different concentrations of DZNep for 7 days. Then the ALP staining was performed after cells were fixed with 4% paraformaldehyde. Scale bar = 20 μm. C Percentage of ALP + area of total area was quantified by ImageJ (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). D The Alizarin Red staining was performed after BMSC were treated with osteoblast-cultured medium and with indicated concentrations of DZNep for 28 days. Scale bar = 50 μm. E Quantification of area of calcium deposition in the extracellular matrix (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). F ALP was detected after BMMs and osteoblast precursor cells (calvarial OB-derived) were co-cultivated in presence or absence of DZNep for 7 days. G Percentage of ALP + area of total area was analyzed by ImageJ (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). Data are expressed as median and interquartile range, n = 3–4