Fig. 3: CFP1-dependent epigenetic regulation intervenes in important uterine P4 responses via H3k4me3-dependent and H3k4me3-independent manners in mice.
a Venn diagram of identifying genes that overlapped in datasets between CFP1-binding sites on their extended gene bodies (2 Kbp upstream and 200 bp downstream of TSS and CGI), downregulated gene in Cfp1d/d mice (>1.5 folds), and downregulated H3K4me3 sites in Cfp1d/d mice (>1.5 folds) on Day 4. b Schematic cartoon to show the epithelium–stroma crosstalk mediated by SSP. c Real-time RT-PCR analyses of upstream and downstream genes in IHH-dependent SSP (Gata2, Sox17, Nr2f2, Hand2, Fgf2, and Fgf18). n = 8 biologically independent samples per genotype. Data are presented as mean values with SD. Statistical analyses were performed using the unpaired Student’s t-tests. *p < 0.05, **p < 0.01. d Integrative Genomics Viewer screenshots that show the distribution of CFP1 binding, H3K4me3 site, and RNA expression intensity in Gata2, Sox17, and Ihh of Cfp1f/f and Cfp1d/d mouse uterus on Day 4. e Real-time ChIP PCR for detecting CFP1, SETD1, and H3K4me3, binding on Gata2, Sox17, and Ihh in the uterus of Cfp1f/f and Cfp1d/d mice on Day 4. n = 3 biologically independent samples per genotype. Data are presented as mean values with SD. Statistical analyses were performed using the unpaired Student’s t-tests. *p < 0.05, **p < 0.01. f Immunofluorescent staining of KI67 in the uterus of the OVX Cfp1f/f and Cfp1d/d mice treated with E2 and/or P4 for 24 h. At 3 h after E2 or E2 + P4, one uterine horn was injected with SAG and the other one with PBS. Green and red indicate KI-67 and the nucleus, respectively. S stroma, GE glandular epithelium, LE luminal epithelium. Scale bar, 50 µm. g Percentages of KI67-positive cells in (f). n = 4 biologically independent samples per genotype. Data are presented as mean values with SD. Statistical analyses were performed using the unpaired Student’s t-tests. *p < 0.05, **p < 0.01. h Real-time RT-PCR analyses of SSP genes (Gli1, Gli2, Nr2f2, and Hand2) 3 h after SAG injection. n = 8 biologically independent samples per genotype. Data are presented as mean values with SD. Statistical analyses were performed using the multiple comparisons. *p < 0.05, **p < 0.01.
