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The bone morphogenetic protein (BMP)‐2 is a potent osteoinductive signal, inducing bone formation in vivo and osteoblast differentiation from non‐osseous cells in vitro. The runt domain‐related protein Cbfa1/PEBP2αA/AML‐3 is a critical component of bone formation in vivo and transcriptional regulator of osteoblast differentiation. To investigate the relationship between the extracellular BMP‐2 signal, Cbfa1, and osteogenesis, we examined expression of Cbfa1 and osteoblastic genes during the BMP‐2 induced osteogenic transdifferentiation of the myoblastic cell line C2C12. BMP‐2 treatment completely blocked myotube formation and transiently induced expression of Cbfa1 and the bone‐related homeodomain protein Msx‐2 concomitant with loss of the myoblast phenotype. While induction of collagen type I and alkaline phosphatase (AP) expression coincided with Cbfa1 expression, Cbfa1 mRNA was …