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Figure 1 Foxf1+ Mesenchyme Promotes Medial Constriction at the Sox2-Nkx2-1 Boundary
(A–C′) Immunostaining showing TE morphogenesis in Xenopus laevis. (A–C) Surface renderings of whole mount confocal images and (A′–C′) transverse optical sections. dorsal foregut; dfg, ventral foregut; vfg, esophagus; e and trachea; t. Scale bar, 100 μm.
(D) Medial constriction in X. laevis with quantification of the difference in foregut width between NF34 and NF35. Scale bar, 100 μm. Difference of means test, *p < 0.05.
(E) Transgenic membrane-GFP X. laevis show increased medial mesenchyme cell density. Scale bar, 100 μm. Student’s two-tailed t test, *p < 0.05.
(F) Medial constriction mouse embryos showing. Dashed yellow lines denote medial mesoderm. Scale bar, 100 μm.
(G) Average change in mesoderm width between E9.5 and E10.0 was not significant. Difference of means test.
(H) Average mesoderm cell density at E9.5 and E10.0 was not significantly different. Student’s two-tailed t test.
(I) Nkx2-1 and Sox2 mouse mutants (E10.5-E11) fail to undergo medial constriction. Scale bar, 100 μm.
(J) Removal of the lateral plate mesoderm prevents medial constriction in X. laevis embryos. Student’s two-tailed t test between side without mesoderm and either control embryos or the contralateral side. *p < 0.05. Scale bar, 100 μm.
(I) Summary of medial constriction. |
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Figure 2 Endosome-Mediated Epithelial Remodeling Is Required for TE Septation
(A) Model of epithelial fusion.
(B and C) Sequential optical sections of X. laevis (B) and mouse (C) embryo immunostaining showing loss of aPKC and increased integrin or Cdh1 at the contact point (arrow, ii). Scale bar, 50 μm.
(D) A unique population of cells co-expressing of Sox2 and Nkx2-1 in the mouse foregut. Scale bar, 100 μm.
(E) Rab11 and Cdh1 are enriched in the X. laevis septum. Scale bar, 50 μm.
(F) Rab11, aPKC, and Cdh1 enriched at the fusion point in mouse. Scale bar, 50 μm.
(G and H) Inhibition of endosome recycling by dynasore treatment of X. laevis (NF32-41) results in a failure to reduce aPKC at NF41 (G) and a TEC at NF44 (H). Scale bar, 100 μm.
(I) Quantification of reduced trachea (t) length relative to the laryngotracheal (ltg) segment in NF42-44 X. laevis embryos. Student’s two-tailed t test, between manipulated and control sibling embryos ∗p < 0.05.
(J) Rab11a CRISPR-mediated mutation or MO knockdown results in a TEC at NF42-44 in X. laevis. Scale bar, 50 μm. |
