Fig. 5: Design of a reversible KEP expression system for in vivo studies.

a Experimental scheme illustrating the generation of KEPi_VAL + S and KEPi_VAL + S-S cell line. b G-banding karyotype analysis of KEPi#28_VAL + S cells. c Western blot analysis of KLF2-V5 and HA-ERAS expression in KEPi#13, KEPi#18, KEPi#28, and KEPi#36 cells, before and after withdrawal of Shield1 (two biological replicates). Source data are provided in the Source Data file. d Two-dimensional PCA of transcriptomic profiles from the indicated cell lines (three biological replicates). e A volcano plot of differentially expressed genes (DEGs) between KEPi_VAL + S and KEPi_VAL + S–S cells, based on a Mann–Whitney U-test. f Percentage of rabbit embryos containing GFP+ cells after microinjection of each cell line (three independent experiments). g Distribution of embryos according to the number of GFP⁺ cells per embryo (three independent experiments). h Distribution of embryos according to the number of GFP⁺/SOX2⁺ cells per embryo (three independent experiments). i Confocal images of late-blastocyst-stage rabbit embryos (E5.0, 3DIV) after microinjection of KEPi_VAL + S and KEPi_VAL + S-S cells into morula-stage (E2.8) embryos. Scale bars: 50 µm. Images are representative of three independent experiments per cell line and condition.