The MEF2 gene family encodes a MADS-box transcription factor which regulates expression of many muscle-specific genes. We examined the expression of the MEF2 genes in mouse embryonal carcinoma P19 cells before and after in vitro muscle differentiation induced by dimethyl sulfoxide (DMSO). At least three different MADS/MEF2 domains (MEF2A, 2B and 2D) were isolated from P19 cells with the MOPAC technique (mixed oligonucleotides primed amplification of cDNA). Although two of the MADS/MEF2 domain sequences were identical to those of human MEF2A and MEF2D, another domain sequence was similar but not identical to that of human MEF2B. While the transcription of MEF2A and MEF2D was up-regulated during differentiation of P19 cells, the MEF2B homologue was abundantly transcribed in undifferentiated P19, F9 and ES cells and down-regulated in adult heart, skeletal muscle or brain, suggesting that the murine MEF2B homologue might have a function distinct from other members of the MEF2 gene family in embryogenesis and development.