Self-renewal of human embryonic stem cells (ESCs) is promoted by truly FGF and TGF beta/Activin signaling, and differentiation is promoted by BMP signaling, but how these signals regulate genes crucial to your servicing of pluripotency is unclear. Making use of a defined medium, we demonstrate here that each TGF beta and FGF signals synergize to inhibit BMP signaling; sustain expression of pluripotency-associated Kinesin genes for instance NANOG, OCT4, and SOX2; and market long-term undifferentiated proliferation of human ESCs. We also demonstrate that each TGF beta- and BMP-responsive SMADs can bind with all the NANOG proximal promoter. NANOG promoter activity is enhanced by TGF beta/Activin and FGF signaling and it is decreased by BMP signaling. Mutation of putative SMAD binding aspects decreases NANOG promoter activity to basal amounts and tends to make NANOG unresponsive to BMP and TGF beta signaling. These success suggest that direct binding of TGF beta/Activin-responsive SMADs for the NANOG promoter plays an crucial part in sustaining human ESC self-renewal.