Fig S1 Mannitol pre-treatment dosen’t affect RANKL-induced macrophage activation. A B To study the effect of mannitol pre-treatment, as a control for osmotic.

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Fig S1 Mannitol pre-treatment dosen’t affect RANKL-induced macrophage activation. A B To study the effect of mannitol pre-treatment, as a control for osmotic pressure, RAW264.7 cells were treated with mannitol or normal glucose condition for 7days, followed by the stimulation with recombinant murine sRANKL. Mannitol pre-treatment did not significantly affect the macrophage activation in both of the number of TRAP positive cells and NFATc1 expression as a master regulator of the RANKL signaling (Fig S1A, B).

Fig S2 Glut 1 and 3 are main Gluts of macrophage. We examined Glut mRNA of macrophages. Glut-1 and 3 were main Gluts of RAW264.7 macrophages.

Fig S3 High glucose pre-treatment suppressed RANKL-induced Akt phosphorylation in macrophage. To examine the effect of recombinant murine sRANKL for phosphorylation of Akt, RAW264.7 cells were treated with high glucose condition (HG group, 15.5 mM glucose) or normal glucose condition (NG group, 5.5 mM glucose) for 7days, followed by the stimulation with recombinant murine sRANKL for 30 minutes, and protein extraction was performed. Murine sRANKL phosphorylates Akt in NG group, but this phosphorylation was suppressed in HG group. (1.53 ± 0.08 vs. 13.07 ± 0.23, P < 0.05) Relative expression levels for western blotting were quantified using the ImageJ program. Values are presented as means ± SD. *P < 0.05.