Fig. 5: N-desulfated heparin with 2-O and 6-O-sulfated groups effectively binds to M-CSF and inhibits osteoclast differentiation. | Cell Death & Disease

Fig. 5: N-desulfated heparin with 2-O and 6-O-sulfated groups effectively binds to M-CSF and inhibits osteoclast differentiation.

From: Suppressive effect of syndecan ectodomains and N-desulfated heparins on osteoclastogenesis via direct binding to macrophage-colony stimulating factor

Fig. 5

a Cell proliferation. Osteoclast precursors (upper panel) and RAW264.7 cells (lower panel) were treated with 2-O, 6-O-, or N-desulfated heparin in the presence and absence of M-CSF for 3 days, respectively. Cell proliferation was determined by MTT assay. b Osteoclast differentiation. Osteoclast precursors were treated with 2-O, 6-O-, or N-desulfated heparin and differentiated into osteoclasts. TRAP-stained cells were photographed (right panel) and TRAP(+) MNCs were counted (left panel). Scale bar, 200 μm. c Gene expression of osteoclastogenic markers. Osteoclast precursors were treated with 2-O, 6-O-, or N-desulfated heparin and differentiated into osteoclasts for 3 days. The expression levels of osteoclast markers were examined by immunoblot analysis. β-Actin was used as a loading control. d M-CSF-specific signaling. M-CSF-induced signals were analyzed as described in Fig. 4d using 2-O, 6-O-, or N-desulfated heparin. e Quantitative binding affinity of M-CSF to syndecan ectodomains was analyzed with a competition assay using 2-O, 6-O-, or N-desulfated heparin as described in Fig. 4e. Results represent the means ± SD (n = 4 as in a; n = 3 as in b and e). The p value indicates the comparison between the treatment group and control. *p < 0.05; **p < 0.01

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