Journal: Journal of Biological Chemistry
Article Title: Stimulation of Bone Formation in Cortical Bone of Mice Treated with a Receptor Activator of Nuclear Factor-κB Ligand (RANKL)-binding Peptide That Possesses Osteoclastogenesis Inhibitory Activity
doi: 10.1074/jbc.m112.426080
Figure Lengend Snippet: FIGURE 4. Participation of BMPs in W9 stimulation in E1 cells. A, E1 cells were cultured in the presence of 50 M W9 or 5 ng/ml BMP-2 with or without 0.25 and 1.0 g/ml sBMPR-1A for 5 days. ALP activity was measured by the pNPP method and expressed as the mean S.D. b, p 0.01 versus W9; d, p 0.01 versus BMP-2. Significant difference was determined using ANOVA with Dunnett’s test. B, postconfluent E1 cells were stimulated with 100 M W9 for 10–360 min, and each whole cell lysate was prepared from the cells using RIPA buffer. Phosphorylated Smad1/5/8, total Smad1, and -actin were detected by Western blotting. C, E1 cells were cultured in the presence or absence of 100 M W9 with or without 0.4 and 2.0 g/ml anti-BMP-2/4 neu- tralizing antibody (BMP-2/4) for 5 days. Data were measured as described above and expressed as the mean S.D. b, p 0.01 versus W9. Significant difference was determined using ANOVA with Dunnett’s test.
Article Snippet: Recombinant human TNF- , recombinant human BMP-2, anti-BMP-2/4-neutralizing polyclonal antibody, recombinant human soluble BMP receptor type 1A (sBMPR1A), recombinant human TNF- , recombinant humanWnt5a, recombinant human Wnt10b, and recombinant mouse Dickkopf-related protein (Dkk-1) were purchased from R&D Systems (Minneapolis, MN).
Techniques: Cell Culture, Activity Assay, Western Blot