Fig. 4: Esrrb controls the epithelial programme via Spry2. | Nature Communications

Fig. 4: Esrrb controls the epithelial programme via Spry2.

From: Wnt/Beta-catenin/Esrrb signalling controls the tissue-scale reorganization and maintenance of the pluripotent lineage during murine embryonic diapause

Fig. 4

a Depletion of Esrrb upon 4OHT treatment. Histone H3 serves as a loading control. Uncropped blots are provided in source data file. b Esrrb fl/fl ES cells expressing tamoxifen-inducible Cre cultured without or in the presence of 4OHT for 1 day before 3D culture. After that, the cells were grown in Matrigel in a medium supplemented with DMSO or CH for 24 h or 48 h and stained for Par6, cleaved Caspase-3 and DAPI. The apical domain in Esrrb-deficient cells is indicated with an arrow. c Establishment of epithelial rosettes in Esrrb Δ/Δ (+4OHT) and control Esrrb fl/fl (−4OHT) ES cells, Data represent mean ± SD, three independent experiments, two-tailed unpaired Student’s t test, the exact P value is noted in the figure. d Scatter plot of gene expression of Esrrb Δ/Δ and Esrrb fl/fl cells grown in the presence of CH for 24 h. Differentially expressed genes that are significantly up- or downregulated are shown in red, adjusted P value < 0.01, three replicates per genotype. e Gene expression of Wnt-responsive naive pluripotency factors in Esrrb Δ/Δ and Esrrb fl/fl cells. f GSEA plots showing enrichment of MAPK, apoptosis, focal adhesion and tight junction KEGG pathways. g Gene expression of epithelial factors in Esrrb Δ/Δ and Esrrb fl/fl cells. h Gene tracks representing the binding of Esrrb at the indicated loci. The x axis represents the linear sequence of genomic DNA, and the y axis represents the total number of mapped reads. i Percentage of E14 ES cells ectopically expressing Krt18, Ntn1, Arl4c or Spry2 that formed Par6-positive polarised rosettes. Data represent mean ± SD, three independent experiments, two-tailed unpaired Student’s t test, the exact P value is noted in the figure. j E14 ES cells (control) and E14 ES cells ectopically expressing Krt18, Ntn1, Arl4c or Spry2 stained for Par6, Nanog and DAPI. k Inducible ectopic expression of Spry2 in E14 ES cells (+Dox), compared to unstimulated (−Dox) control, stained for Par6, E-cadherin and DAPI. Scale bars, 10 µm.

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