Expression and subcellular localization of ZAS3 associate with osteoclastogenesis of RAW 264.7 preosteoclasts.

(A) Morphological changes and increase in TRAP expression of RAW264.7 cells upon sRANKL-mediated osteoclastogenesis. RAW 264.7 cells were cultured for 0, 2, 4 or 6 days and stained for TRAP. Scale bar, 50 µm and arrows show representative TRAP+ cells. (B) Western blot analysis to show the presence of ZAS3 in RAW 264.7 cells at days 0, 2, 4 and 6 incubated with sRANKL (50 ng/ml). The protein filter was also incubated with histone H1 antibodies as a loading control. (C) Immunohistochemical fluorescence microscopy showing the expression of ZAS3 in RAW 264.7 cells at various stages of osteoclast differentiation. Number shown was the number of nucleus or nuclei presence in the cell indicated. During osteoclastogenesis, multinucleated giant cells are progressively formed by cell fusion. Therefore, increasing number of nuclei represented cells were at more advanced stage of osteoclastogenesis.