Fig. 4: Procedures for decidualization deregulated by uterine Men1 loss.

a Volcano plot of differentially expressed genes in Men1^f/f and Men1^d/d decidual tissues on day 8 as determined by RNA-seq. Blue and red dots represent genes that are differentially expressed (p < 0.05) between Men1^f/f and Men1^d/d decidual tissues by exact negative binomial test in edgeR package of R. Genes not significantly differentially expressed are shown in black. b Gene Ontology functional analysis for differentially expressed genes determined by DAVID by Kappa Statistics (p < 0.05). n number of genes. c Heatmap of physiological function associated genes differentially regulated between Men1^f/f and Men1^d/d uteri on day 8. RPKM of each gene from different samples was normalized to Z-score. d In situ hybridization of Ptn and Sfrp4 in Men1^f/f and Men1^d/d implantation sites on day 8. Scale bar: 200 μm. e Quantitative real-time PCR analysis of Angpt1 and Angpt2 mRNA levels in Men1^f/f and Men1^d/d implantation sites on day 8. The values are normalized to Gapdh and indicated as the mean ± SEM (n = 3 biologically independent samples). Two-tailed unpaired Student’s t-test, *p = 0.025 (Angpt1), *p = 0.039 (Angpt2). f Immunohistochemical analysis of p27 in the SDZ of Men1^f/f and Men1^d/d implantation sites on day 8. Scale bar: 200 μm. g GSEA plot showing the enrichment of cytokinesis-related genes (n = 115) in Men1^d/d uteri compared with Men1^f/f uteri on day 8. NES normalized enrichment score. ***p = 0.001. h Heatmap of cytokinesis associated genes differentially expressed between Men1^f/f and Men1^d/d mice uteri on day 8. RPKM of each gene from different samples was normalized to Z-score. i Immunohistochemical analysis of pH3 in Men1^f/f and Men1^d/d implantation sites on day 8. Scale bars: 200 μm. j Quantitative analyses of pH3 positively stained cells in the SDZ on day 8. Data represent the mean ± SEM (n = 4 biologically independent samples). Two-tailed unpaired Student’s t-test, **p = 0.0051.