Fig. 5. Regulation of RANKL-activated signaling pathways by SPA0355.

(A) BMMs were treated with M-CSF (10 ng/ml) and sRANKL (30 ng/ml) with or without 10 µM SPA0355 for the indicated periods. Cell lysates were prepared, and the changes of MAPKs, Akt, and NF-κB were evaluated by western blotting. HSP90 and lamin B were used as loading controls for cytosolic and nuclear protein, respectively. (B) BMMs were transiently transfected with an NfkB luciferase construct and then treated with 30 ng/ml RANKL in the presence or absence of 10 µM SPA0355. After 24 h, cells were harvested in reporter lysis buffer, and luciferase activity in cell lysates was assayed (n=4). Values are the mean±SEM. ^*p<0.05 vs. untreated control. ^#p<0.05 vs. RANKL+VEH.