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. Author manuscript; available in PMC: 2020 Oct 1.
Published in final edited form as: J Orthop Res. 2019 Jun 24;37(10):2089–2103. doi: 10.1002/jor.24384

Figure 3. Genetic inhibition of Notch signaling suppresses osteoclast maturation but does not impact early gene expression.

Figure 3.

Bone marrow-derived macrophages were collected from both WT and dnMAML mice and differentiated into osteoclasts ex vivo. (A) TRAP stained osteoclasts (purple) in WT group were giant and well-fused while osteoclasts from Notch-inhibited mice failed to fuse effectively and remained immature. (B) Quantification of TRAP stained multinuclear osteoclasts showed a significant reduction in average number of osteoclasts in dnMAML group. (C) To delineate cytoskeletal reorganization, actin filaments were assessed using Phalloidin staining. There were fewer and smaller osteoclasts in dnMAML group, but these cells still demonstrated podosome belts, suggesting normal actin organization. (D) and (E) Expression analysis of osteoclastic marker genes (TRAP, CTSK, MMP9 and Wnt10b) and osteoclast specific fusion markers (DC-STAMP and OC-STAMP) ex vivo showed no significant differences between the dnMAML and WT groups. Values represented are Mean ± S.D from at least three independent experiments. P value was calculated using Student t-test (*P<0.05).