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. 2020 Mar 19;117(14):7755–7763. doi: 10.1073/pnas.1915582117

Fig. 3.

Fig. 3.

AHCY T136 O-GlcNAcylation promotes mESC pluripotency and self-renewal. (A) Immunoblotting of cellular O-GlcNAcylation levels in E14.1 cells upon induction of differentiation. (B) Immunoblotting of AHCY and its glycosylation levels during the first 2 d of cell differentiation (n = 3 assays). (C) Enzymatic activity of AHCY at indicated time points of cell differentiation (n = 3 assays). (D) The number of colonies staining positive for AP produced from AHCY WT or T136A rescue E14.1 cells (n = 3 assays). (E) Relative clone numbers produced from AHCY WT or T136A rescue E14.1 cells (n = 3 assays). (F) Teratomas formation in nude mice (n = 5 per group) injected with AHCY WT or T136A rescue E14.1 cells. (G) Relative expression levels of the differentiation markers in teratoma derived from WT or T136A AHCY rescue E14.1 cells (n = 3 assays). (H) Relative abundance of metabolites in the methionine cycle in the WT or T136A AHCY rescue E14.1 cells (n = 3 assays). (I) Immunoblotting and quantification of trimethylation of histone H3 lysine residues in AHCY WT or T136A rescue E14.1 cells (n = 3 assays). (J) Expression profiles of selected genes involved in cell stemness and differentiation. Error bars denote the means ± SEM. Statistical analyses were performed by unpaired Student’s t test (*P < 0.05, **P < 0.01).