Fig. 1: Ectodomains of syndecan-1 to 4 from HEK293E cells block osteoclast differentiation. | Cell Death & Disease

Fig. 1: Ectodomains of syndecan-1 to 4 from HEK293E cells block osteoclast differentiation.

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

Fig. 1

a Schematic diagram of mouse syndecan-1 to 4 ectodomains (mSDC1ED to mSDC4ED) expressed from HEK293E cells. The gray and black boxes represent the syndecan ectodomain and histidine (His), respectively (left panel). The recombinant syndecan ectodomains purified from HEK293E cells were separated by electrophoresis on a 10% SDS-PAGE and immunoblotted with anti-His antibody (right panel). The positions of the molecular mass markers are represented on the left. b Inhibitory effect of syndecan ectodomains on osteoclast differentiation. Bone marrow-derived osteoclast precursors were treated with various concentrations of recombinant syndecan ectodomains in the presence of M-CSF (30 ng/mL) and RANKL (100 ng/mL) for 4 days. Cells were fixed and stained for TRAP. The number of TRAP-positive osteoclasts [TRAP(+) MNCs] was counted under a light microscope. Scale bar, 200 μm. c Osteoclastogenic marker genes. Osteoclast precursors were treated with syndecan ectodomains (1 nM) in the presence of M-CSF and RANKL for 4 days. Cell lysates were subjected to immunoblot analysis using antibodies specific for NFATc1, c-Fos, and ATP6V0D2. β-Actin was used as a loading control. Asterisk indicates non-specific bands. d Osteoclast precursors were treated with syndecan ectodomains (1 nM) at the indicated time points during osteoclast differentiation. After TRAP staining, the number of TRAP(+) MNCs was counted. Results represent the means ± SD (n = 3). **p < 0.01

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