Fig. 5: Ex vivo uterus replicates robust induction of COX-2 and mirrors decreased trophoblast invasion by COX-2 inhibition. | Nature Communications

Fig. 5: Ex vivo uterus replicates robust induction of COX-2 and mirrors decreased trophoblast invasion by COX-2 inhibition.

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

Fig. 5

A, B Gene ontology analysis of uterine scRNA-seq found enriched lipid metabolic pathways in the attached stroma clustered in Fig. 3C. Modified Fisher’s exact test with P value cutoff   0.05 was used (A). C Attached stroma was detected in in vivo implantation site (IS) and ex vivo inter-implantation site (I-IS) and IS. D Ptgs2 expression in ex vivo attached stroma was highly induced in IS (attached by embryo) compared to I-IS (attached by PDMS), suggesting robust signals from embryos. The P-value was calculated by two-tailed Mann-Whitney’s U-test. E Immunofluorescence of COX-2 on histological sections detects localized expression of COX-2 exclusively at the attachment sites of the endometria both in vivo and ex vivo. F-K. A COX-2 inhibitor celecoxib retards embryonic attachment and trophoblast invasion. Each graph represents the percentage of embryos that attached at 24 h/initially loaded (F), shed between 24 h and 48 h/attached at 24 h (G, H), and sustained on the endometria at 48 h/initially loaded (I). Mean ± SEM (F, G, I). A stereomicroscopic photo (H) shows spontaneously shed embryos at 48 h indicated by arrows. Scale bar: 500 μm. Slices from three-dimensional images show representative trophoblast invasion (J). Examples of higher and lower invasions in each group were presented and the invasion volume was compared (K). The box plot is presented as minimum, lower quartile, median, upper quartile, and maximum. + represents the mean value. White and orange dotted lines represent the surface of LE and the boundary of LE and Str, respectively. Two-tailed unpaired Student’s t-test (F, G, I, K) was used (n = 6 vs 7 (F), 4 vs 5 (G), 4 vs 5 (I), 8 vs 9 (K) biologically independent samples). *, P < 0.05; **, P < 0.01; ***, P < 0.001. White and orange dotted lines represent the surface of LE and the boundary of LE and Str, respectively. DsRed2 or tdTomato indicates the mitochondria or cell membrane of all embryonic/extraembryonic cell lineages. Scale bars: 50 μm. Each plot represents the result per a recipient mouse (F, G, I) or the value for an embryo (K).

Back to article page