Fig. 3: Gm26793-null embryos exhibit developmental abnormalities during the lineage segregation between epiblast and primitive endoderm.

a Experimental overview of mouse breeding, embryo preparation and the functional assays conducted at the indicated developmental stages. b Representative images showing the resorbed decidua in GKO mouse at E7.5, which were highlighted by the arrows. c Quantification of the normal decidua collected at E7.5 and the acquired offspring number per litter. N represents the number of mating females. d, e Representative images showing the embryos collected at early (E3.5) (d) and late (E4.5) (e) blastocyst stage for both WT and GKO groups. Abnormal embryos were indicated by red arrows. f Quantification of the normal developmental rates at early and late blastocyst stage. N represents the total number of blastocysts collected. Data are summarized from three biologically independent experiments. g UMAP plots of scRNA-seq data from E3.5 and E4.5 blastocysts, respectively. The percentages of cell type composition are highlighted as a pie chart embedded in the related UMAP plot. Degenerative cells are circled by the oval. h Pseudotime trajectories of pre-implantation cell lineages inferred by Monocle 3 (left). Visualization of differentiated and transient epiblast cells in UMAP plots based on the trajectory branch towards primitive endoderm (right). The percentages of epiblast subtype cells are highlighted as a pie chart embedded in the related UMAP plot. i Heatmap showing DEG groups among different epiblast subtypes displayed in h. Representative genes are listed on the right panel. j Box plots showing average expression levels of differentiated and transient epiblast-specific genes in the indicated cell clusters. k Antagonistic distribution and expression level of primitive endoderm and epiblast marker genes among transient and differentiated epiblast cells. l Heatmap showing the transcriptomic correlation within in vitro EBs, differentiated and transient epiblast. All data are shown as means ± SEM. Student’s t-test analysis is used in c, f; ***P < 0.001.