Fig. 3: The Pten-inhibitor SF1670 contributes to sustaining ESC pluripotency.

a Phase-contrast images of WT ESCs treated with DMSO and the Pten-inhibitor SF1670 when cultured in medium with 2i and LIF (2iL). Scale bars, 100 µm. b Pten-inhibitor SF1670 promotes the ESC pluripotency. c Alkaline phosphatase staining of WT ESC colonies treated with DMSO and SF1670. There are more flattened colonies (black arrow) in DMSO-treated ESCs, and more domed colonies (green arrow) in SF1670-treated ESCs. d Analysis of colony morphology of WT ESCs treated with DMSO and SF1670. Error bars indicate mean ± SEM (n = 3), and 80 colonies were scored in each replicate. e qRT-PCR analysis of mRNA expression of pluripotency markers (Esrrb, Zfp42, Fgf4, Nanog, Oct4, Rexo1, Dppa3, and Lefty2) in DMSO-treated ESCs and SF1670-treated ESCs. Error bars indicate mean ± SD (n = 3). f Western blot analysis of WT ESCs treated with DMSO and SF1670 showing the expression of the pluripotency markers Nanog, Klf4, and Oct4. g Phase-contrast images of DMSO-treated WT, SF1670-treated WT, and Pten^−/− ESCs when cultured in medium with 2iL. Scale bars, 100 µm. h AP staining of DMSO-treated WT, SF1670-treated WT, and Pten^−/− ESC colonies when cultured in medium with 2iL. i Analysis of colony morphology of DMSO-treated WT, SF1670-treated WT, and Pten^−/− ESC colonies. Error bars indicate mean ± SEM (n = 3), and 80 colonies were scored in each replicate. j Nude mice were injected with WT ESCs, Pten^−/− ESCs, and SF1670-treated WT ESCs on the left and right sides, respectively. Teratomas were generated at the contralateral positions aligned at the same positions in each treatment. k Weights (g) of teratomas from WT ESCs, Pten^−/− ESCs, and SF1670-treated WT ESCs.