Fig. 6

Activation of Notch reversed the negative effects of M1 macrophages on osteocyte morphology and maturation. A(i) SEM images showed osteocyte morphology changed after activation of Notch signaling pathway (a, c and e: Low magnification at ×500 and the scale bars represented 20 μm; b, d and f: High magnification at ×5000 and the scale bars represented 5 μm); A(ii) The bar graphs showed the morphology changes of osteocytes after 14 days simulation with macrophage-derived conditioned medium with or without activation of Notch signaling pathway. A total of 20 randomly selected areas from each sample were analyzed, and data shown as the mean ± SD (*p < 0.05, one-way ANOVA); B(i) epi-fluorescence images of M1 macrophage-stimulated osteocytes after Notch activation; B(ii) the intensity of DMP-GFP was measured using ImageJ software; C gene expression of osteocyte markers after 14 days simulation with macrophage-derived conditioned medium, with or without activation of Notch signaling pathway. Data from three independent experiments performed under the same condition were shown as mean ± SD (*p < 0.05, one-way ANOVA)