Fig. 7: Embryonic AKT1 transduction ameliorates invasion defects in the uteri with celecoxib administration ex vivo and in vivo. | Nature Communications

Fig. 7: Embryonic AKT1 transduction ameliorates invasion defects in the uteri with celecoxib administration ex vivo and in vivo.

From: An ex vivo uterine system captures implantation, embryogenesis, and trophoblast invasion via maternal-embryonic signaling

Fig. 7

A–E Ex vivo experiments. A schematic diagram of the ex vivo rescue for celecoxib-induced invasion defect using myr-AKT1-AAV1. AT, acidified Tyrode’s solution. B Attachment at 24 h/initially loaded. C. Sustention on the endometria at 48 h/initially loaded. Mean ± SEM (B, C). D Representative slices from three-dimensional images. E. Quantitative data of the invading volume. The box plot is presented as minimum, lower quartile, median, upper quartile, and maximum. + represents the mean value. F–J In vivo experiments. F A schematic diagram of the in vivo rescue for celecoxib-induced implantation defect using myr-AKT1-AAV1. G The uteri at dpc 5.5 after intravenous injection of blue dye solution. Implantation sites (ISs) are indicated by arrows. H Number of IS per mouse. 16 embryos were transferred to each pseudopregnant mice. Mean ± SEM. I The weight of IS. The box plot is presented as minimum, lower quartile, median, upper quartile, and maximum. + represents the mean value. J Representative photos of immunofluorescence for CK8 on histological sections. Trophoblast invasion was limited in celecoxib+EGFP-AAV1 group compared to vehicle+EGFP-AAV1 and celecoxib+myr-AKT1-AAV1 groups. Yellow arrowheads represent embryos. White dotted line indicates the outer border of invading trophoblasts. GE, glandular epithelium; Str, stroma; Tr, trophoblast. Ordinary one-way ANOVA with post hoc Tukey’s multiple comparisons test (B, C, E, H, I) was used (n = 5 vs 5 vs 5 (B, C), 13 vs 10 vs 8 (E), 4 vs 6 vs 7 (H), 57 vs 16 vs 46 (I) biologically independent samples). *, P < 0.05; **, P < 0.01; ****, P < 0.0001; ns, not significant. tdTomato indicates the cell membrane of all embryonic/extraembryonic cell lineages. Scale bars: 50 μm. K Proposed models of how uterine COX-2 activates embryonic AKT. In model A, PGs bind to PG receptors in TE and activate AKT. In model B, cytokines and growth factors produced downstream of endometrial PG receptors accelerate AKT phosphorylation. Each plot represents the result per a recipient mouse (B, C, H) or the value for an embryo (E, I). A, F, and K created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en).

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