Research Paper Volume 12, Issue 6 pp 4727—4741

Metformin coordinates osteoblast/osteoclast differentiation associated with ischemic osteonecrosis

Figure 3. Increase in phosphorylation of p38 by metformin. (A) Cells were treated with 2.5, 5, and 10 μM of metformin or an equal volume of DMSO (0.1%) for 1 h. Cells were harvested, and the lysed proteins were resolved on SDS-PAGE and immunoblotted with specific antibodies against p-p38 MAPK and p38 MAPK. (B) For immunoprecipitation analysis, cells were treated with 5 μM of metformin or an equal volume of DMSO (0.1%) for 1 h. Cells were harvested, and the lysed proteins were immunoprecipitated with antibody against p38 MAPK. The immunoprecipitated proteins were resolved on SDS-PAGE and immunoblotted with specific antibodies against Runx2, OSX, p-p38 MAPK, and p38 MAPK. IgG heavy chain blotting was used as a loading control.