Fig. 2

Characteristics of ciXEN cells at passage 5. Morphological appearances of ciXEN cells at low density and high density (bar, 100 μm) (a). qPCR results for the expression of XEN-related genes (Foxa2, Sox17, Gata4, and Gata6) (b), pluripotency markers (Sox2, Oct4, and Nanog) (c), fibroblast markers (Pdgfrb, Prrx1, Thy1, and Col1a1) (d), and epithelial-related genes (e). f, g Immunofluorescence of Sox2, Gata4, E-cadherin, vimentin, Oct4, Nanog, Sox17, and Foxa2 in ciXEN cells (bar, 50 μm). h Western blot analysis for the expression of Foxa2, Sox17, Gata4, Sox2, E-cadherin, and vimentin in MEFs and ciXEN cells. i EdU assay for proliferative ability of MEFs and ciXEN cells at p5, p10, and p30 (bar, 50 μm). j Detection of cell cycle by flow cytometry. The percentage of G1-, S-, and G2-phase in the cell cycle and the PI of MEFs and ciXEN cells at p5, p10, and p30. p5, passage 5; p10, passage 10; p30, passage 30. *p < 0.05, **p < 0.01, ***p < 0.001. Mean ± SEM, t test and two-way ANOVA, n = 3