Non-coding RNAs (ncRNAs), such as microRNAs and large intergenic non-coding RNAs, have been shown to play essential roles in regulating pluripotency. Yet, it is not clear the role of natural antisense transcripts (NATs), also belonging to ncRNAs, in embryonic stem cells. However, the role of NATs in embryonic stem cells remains unknown. We further confirmed the expression of the NATs of three key pluripotency genes, Oct4, Nanog and Sox2. Moreover, overexpression of Sox2-NAT reduces the expression of Sox2 protein, and slightly enhances the Sox2 mRNA level. Altogether, our data indicated that like other ncRNAs, NATs might be involved in pluripotency maintenance.