Fig. 1.

Rapamycin inhibits Epo-dependent osteoblast differentiation in hBMSC. hBMSC were cultured in differentiation inducing media containing Epo and/or rapamycin and assayed via alkaline phosphatase staining after 2 weeks (A) and Alizarin red staining after 3 weeks (B). Staining was quantified using Image J software and normalized to cell number. The experiments were performed twice. (C) Protein extracts from hBMSC cells were analyzed by Western blotting using antibodies against the indicated proteins. (D) hBMSC cells were treated for 24 hours with/without Epo. Expression of EpoR was quantified by qPCR. The results were normalized to β-actin. The results represent average values from triplicate assays and the experiments were performed twice.