The GATA family of transcription factors are implicated in early embryonic development. There are six factors in this family in vertebrates. GATA4 and GATA6 have been demonstrated to induce mouse embryonic stem (mES) cells differentiation toward extraembryonic endoderm (ExE). We investigated the effect of GATA3 on the differentiation of mES cells both in the ES cell and in the embryoid body (EB) states. The results demonstrate that GATA3 overexpression can initiate the ES cell differentiation program toward ExE. Furthermore, overexpression of GATA1 and GATA2 in ES cells and EBs resulted in similar effects. We believe this finding can augment our understanding of mouse ES cell differentiation.