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. 2017 Jul 19;14(9):840–852. doi: 10.7150/ijms.19268

Figure 7.

Figure 7

Effect of Niclosamide on osteoblast differentiation of MC3T3-E1 cells. Cells were treated with the different concentration of niclosamide in the presence Osteoblast differentiation medium for 7 or 21 days, and then the effects of the niclosamide on ALP activity and ECM calcification were evaluated by the ALP or Alizarin red S staining methods. (A) ALP staining on day 7. (B) Alizaring red S staining on day 21. (C) ALP activity on day 7. (D) ECM calcification was quantified by the amount of cell-bound Alizarin red S. Alizarin red S was released by incubation in 10% cetylpyridinium chloride and then detected by measuring the absorbance at 540 nm. ECM calcification was determined relative to the DMSO control. Data are presented as the mean ± SD of three independent experiments. (E) Gene expression levels of RUNX-2, ALP, Col I and OCN on day 7 were determined by real-time PCR analysis. The data were normalized to GPADH. * P < 0.01 and ***P < 0.001, versus the DMSO control.