Fig. 1

AKT induces Nanog promoter in a SUMOylation-dependent manner in PSCs. A Schematic diagram of the experimental design. B iPSCs were transfected with either an AKT1 variant (indicated under each bar) or the empty vector (basal), along with the Nanog5P luciferase reporter. A vector encoding the dominant negative UBC9(C93S) was included (+ , dot-patterned bars) or not (−), as indicated. C, D ESCs were cultured in standard conditions (+ LIF/2i) or were induced to differentiate by LIF/2i withdrawal for 48 h, and then fixed for immunostaining against NANOG. Representative images of phase-contrast microscopy (C) and epifluorescence microscopy (D). Nuclei were stained with DAPI. E Schematic diagram of the experimental design for luciferase experiments at early differentiation. F, G ESCs were transfected with either an AKT1 variant or the empty vector (basal), along with the Nanog5P luciferase reporter and cultured for 48 h in -LIF/2i medium. H U2-Os cells transfected with the vector indicated under each bar, along with the Nanog5P luciferase reporter. In all cases, results were referred to the corresponding basal of each condition (B: dashed line, F: + LIF/2i, G and H: basal) and are shown as mean ± SEM of three independent experiments. When corresponding, different letters indicate significant differences among treatments (p < 0.05), asterisks indicate significant differences of each condition compared to the basal (p < 0.05), and hashes indicate significant differences between fold change of the same AKT1 variant in the experiments comparing with ( +) and without (-) Ubc9(C93S) (p < 0.05). Scale bars: 100 µm