Figure 6.

Effect of seladin-1 small interfering (si) RNA on lipopolysaccharide (LPS) -induced osteoclast formation and alkaline phosphatase (ALP) activity. RAW 264.7 cells were transfected with seladin-1 siRNA or negative control siRNA for 48 hr and the transfected cells were stimulated with LPS (100 ng/ml) for 48 hr. (a) The frequency of osteoclasts was determined with tartrate-resistant acid phosphatase staining. (b) ALP activity was measured using an enzyme assay using p-nitrophenol as the substrate. *P <0·01 versus control siRNA. (c) RAW 264.7 cells were stimulated with receptor activator of nuclear factor-κΒ ligand (RANKL; 100 ng/ml) and macrophage colony-stimulating factor (M-CSF; 25 ng/ml) for various lengths of time. The expression of seladin-1 protein was determined with immunoblotting (d). A typical experiment of three independent experiments is shown.