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Abstract
O-GlcNAcylation is one of the major forms of protein glycosylation and also an important post-translational modification discovered in recent years. To study the roles of O-GlcNAcylation in human pluripotent stem cells we carried out O-GlcNAc transferase gene knock-down and O-GlcNAcase inhibition assays to manipulate global O-GlcNAcylation level in human pluripotent stem cells, splanchnic mesoderm cells, definitive endoderm cells and neural progenitor cells. These experiments indicated that loss of high global O-GlcNAcylation in human pluripotent stem cells 1) elevates the expression of early mesoderm and early endoderm markers such as T, GSC and EOMES, 2) increases their doubling time, elongates G1 phase and shortens S and G2/M phases, however, 3) it doesnt affect differentiation to neural progenitor cells. 4) OGT knockdown during differentiation towards splanchnic mesoderm and definitive endoderm cells elevated their marker gene expression. 5) Enhanced O-GlcNAcylation impairs transcriptional expression of early mesoderm marker genes. The above findings serve to facilitate our understanding about the role of O-GlcNAcylation in human pluripotent stem cells.