Fig. 5: Pten activation by S380A, T382A, T383A mutations suppresses ESC pluripotency.

a, b The domain structure of Pten in the “closed” conformation and “opened” conformation. c Western blots analysis showing the loss of phosphorylation in the Pten-A3 mutant ESCs. d Western blot analysis of WT, Pten-A3 mutant, and Pten^−/− ESCs showing the expression of the pluripotency markers Nanog, Klf4, and Oct4. e Phosphorylation of Akt at S473 and T308 and phosphorylation of Gsk3β at S9 by Western blot. f Scatter plots of transcript expression in WT and Pten-A3 mutant ESCs. Expression values are shown on a log10 scale. Yellow dots indicate upregulated genes in Pten^−/− ESCs, and blue dots indicate downregulated genes (fold-change cutoff = 1.5, FDR threshold ≤ 0.05). g Heat map of FPKM values of pluripotency genes in WT and Pten-A3 mutant ESCs. The heat map was normalized with sigma-normalization per row. See also Table S3. h A nude mouse injected with WT and Pten-A3 mutant ESCs on the left and right sides, respectively. i WT and Pten-A3 mutant ESCs were injected into immunodeficient mice and produced teratocarcinomas containing tissues representative of the three germ layers (mesoderm, ectoderm, and endoderm). j Teratomas from WT and Pten-A3 mutant cells generated at the contralateral positions aligned at the same positions in each genotype. k Weights (g) of teratomas from WT and Pten-A3 mutant cells.