Extended Data Fig. 5: Characterization of NODAL signaling across mouse implantation.
From: Distinct pathways drive anterior hypoblast specification in the implanting human embryo
a, Mean ± S.E.M. nuclear-to-cytoplasmic ratio of total SMAD2.3 within each lineage of segregating and segregated blastocysts, day 7, and day 9 embryos, separated by individual embryo. Note consistency of trends, particularly of TE versus ICM enrichment in segregating blastocysts, and of hypoblast versus epiblast. b, Immunofluorescence images of mouse embryos at E3.5 (N = 10 embryos), E4.5 (N = 6 embryos), E5.5 (N = 6 embryos) and E5.75 (N = 6 embryos) stained for total Smad2.3, DAPI, and either Sox2 or Gata6. c-f, Quantification of nuclear and cytoplasmic Smad2.3. fluorescence and their ratio. g-i, Quantification of the nuclear/cytoplasmic ratio of Smad2.3 within the epiblast (g; n = 63 E3.5, 50 E4.5, 95 E5.5, 95 E5.75 cells), primitive endoderm/visceral endoderm (PrE/VE; n = 69 E3.5, 42 E4.5, 105 E5.5, 129 E5.75 cells) (h) or trophectoderm/extraembryonic ectoderm (TE/ExE) (i; n = 62 E3.5, 60 E4.5, 89 E5.5, 69 E5.75 cells) over time. For Violin plots c-f, central dotted line denotes median and dotted lines mark the 25th and 75th quartiles. Statistical tests: (c) two-sided unpaired T-test; (d-f) Kruskal Wallis with Dunn’s post-hoc; ****p < 0.0001, ***p < 0.001, *p < 0.05. Unmarked pairwise comparisons are not significant (ns). Exact p-values presented in Supplemental Table 8. Scale bars: 100 µm.
