Research Paper Volume 12, Issue 6 pp 4727—4741

Metformin coordinates osteoblast/osteoclast differentiation associated with ischemic osteonecrosis

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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.