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. Author manuscript; available in PMC: 2023 Jun 26.
Published in final edited form as: Dev Cell. 2019 Dec 16;51(6):698–712.e8. doi: 10.1016/j.devcel.2019.11.014

Figure 6. EPS-Blastoids Initiate Implantation In Vivo.

Figure 6.

(A) Number of deciduae obtained after transfer to pseudopregnant females at E2.5 or E3.5. Representative deciduae dissected 4 days post-transfer. NB, natural blastocyst induced; SB, EPS-blastoid induced; M, mesometrial; AM, anti-mesometrial.

(B) Decidua remodelling in natural embryo implantation. S, stroma; G, glands; U, uterine tissue; LE, luminal epithelium; rLE, regressed LE; E, implanted embryo; TGC, trophoblast giant cells; PDZ, primary decidua zone; SDZ, secondary decidua zone.

(C and D) Immunohistochemical staining of the EPS-blastoid-induced decidua. Ptgs2 marks PDZ (C), Ki67 marks normal tissue proliferation of SDZ (D); Yellow asterisk, implantation site; n = 8 SB-induced decidua.

(E and F) Immunohistochemical staining of the decidua at 5.5 d.p.c. Ptgs2 marks PDZ. E, embryo (E). Ki67 marks normal tissue proliferation of SDZ. Yellow asterisk: implantation site (F). n = 2 NB-induced decidua.

(G) Proliferin (PLF) and Krt18 mark TCGs upon invasion. Yellow or black asterisk, implantation site. n = 4 EPS-blastoid-induced decidua.

(H) The EPS-blastoid-induced decidua section shows immunohistoflourescence for Ptgs2 (yellow) at implantation site, Cdx2 (red) for implanting conceptus. Implanted structure magnified, Cdx2 co-localises with ubiquitous TS:eGFP expression (yellow arrows); surrounding cells are positive for nuclear PDGFRa-H2B-GFP (white arrowheads). Right, representative EPS-blastoid before transfer (nuclear PDGFRa-H2B-GFP for PE-like cells and ubiquitous eGFP for TSCs). n = 3 SB-induced decidua.

(I) Break-down of LE during implantation. Top, E-cadherin marks LE before the closure (left); implantation site with lost LE (right). Cdx2 indicates implanted conceptus. Note disrupted structural integrity indicating resorption. Bottom, shows same events during natural implantation (e, embryo).

(J) Immunohistoflourescence for laminin (red) at basement membrane surrounding implanted conceptus (E). Laminin assembly absent in EPS-blastoid-induced deciduae (bottom) n = 3 NB-induced; 5 SB-induced deciduae.

Scale bars represent 20 μm; Error bars represent SEM in all panels.