Nkx2-5 regulates the transcription of muscle-specific genes during cardiomyogenesis. Nkx2-5 expression can induce cardiomyogenesis in aggregated P19 cells but not in monolayer cultures. In order to investigate the mechanism by which cellular aggregation regulates Nkx2-5 function, we examined the role of bone morphogenetic protein 4 (BMP4). We showed that the expression of the BMP inhibitor, noggin, was sufficient to inhibit the induction of cardiomyogenesis by Nkx2-5 during cellular aggregation. Furthermore, soluble BMP4 could activate Nkx2-5 function in monolayer cultures, resulting in the formation of cardiomyocytes. Therefore, BMP signaling is necessary and sufficient for the regulation of Nkx2-5 activity during cardiomyogenesis in P19 cells.