Fig. 3

Characterization of pluripotent identity and competence of a corrected hiPSC clone. A Immunofluorescent staining for NANOG (green) and OCT4 (red) in hiPSCs-c.5483G > A (left) and hiPSCs-cl1 (right). Nuclei were stained with HOECHST (blue). B Flow cytometric analysis of membrane marker TRA-1–81, SSEA-4 and EpCAM in hiPSCs-c.5483G > A (top) and hiPSCs-cl1 (bottom). C Immunofluorescence staining showing the expression of marker genes belonging to the three germ layers in EBs obtained from hiPSCs-cl1. βIII-Tubulin (green), αSMA (red) and GATA4 (red). Nuclei were stained with HOECHST (blue). D qPCR analysis of the three germ layers markers nestin (ectoderm), αSMA (mesoderm) and AFP (endoderm) in EBs obtained from hiPSCs-cl1. EBs derived from commercial hiPSCs were used as positive control (CTR +). Data are means ± SD. Statistical analysis was performed using ordinary one-way ANOVA; ns P > 0.05, ***P ≤ 0.001, ****P ≤ 0.0001