Fig. 7

Implantation initiation of the ETX-embryoids. a Representative implantation sites (IS) in uteri visualized via blue dye after transfer of wild-type embryos and different embryoids. Scale bar, 2 mm. b Percentages of IS per uterus transferred with different embryoids. Two-tailed Student’s t-test, n = 5 mice per group and each recipient transferred with 20 embryoids, three experiments. *P = 0.0193, ***P = 0.0003. c Deciduae volume of E6.5 embryos and different embryoids after transplantation. n = 10 wild-type embryos, 131 ETX-embryoids, 18 ETS-embryoids, 17 EXE-embryoids. d Representative deciduae of wild-type embryos and different embryoids. Scale bar: black, 500 μm; white, 200 μm. e Detection of Dtprp in deciduae of wild-type embryos and different embryoids via in situ hybridization. n = 5 for each group. Scale bar, 500 μm. f, g Representative cross-section by H.E. staining of E5.5 embryo and ETX-embryoid deciduae. n = 4 wild-type embryos and 6 ETX-embryoids. Scale bar, left panel, 500 µm, right panel, 50 µm. h–j Immunostaining of E6.5 embryos and ETX-embryoids’ IS. h, i CK (red). The yellow dotted line shows the uterine axis M-AM. M, mesometrial pole; AM, antimesometrial pole. Scale bar, left panel, 500 µm, right panel, 50 µm. j COX2 (red) and PL1 (green). Bar, 500 µm. n = 5 wild-type embryos, 5 ETX-embryoids. k Immunostaining of CK (red) and PL1 (green) in ETX-embryoids’ IS. Differentiated PL1 + TGCs directly connect with stromal cells of the uterus. White boxes show PL1 and CK-positive cells. Bar, 50 μm. Bar in zoomed, 10 μm. l Immunostaining of Gata4 (red) and Laminin (green) in E5.5 embryos from f and ETX-embryoids’ IS from g. n = 5 wild-type embryos, three ETX-embryoids. Scale bar, 50 µm. m Ratios of embryos with different morphologies detected in the deciduae deduced by E5.5, ETX-, EXE-, and ETS-embryoids. n = 10 deciduae in wild-type, EXE, and ETS-embryoids, 20 in ETX-embryoids. n Morphological analysis of ETX-embryoid in deciduae. “Regular” indicates the ETX-embryoid morphology similar to the wild-type embryos. “Degenerated” indicates the ETX-embryoid degenerated. “None” indicates ETX-embryoid components wasn't detected. n = 20 deciduaes for each stage. Error bar are means ± s.e.m. Source Data of b, c, m, n are provided as the Source Data file