Journal: Nature cell biology

Article Title: Elevated contractility drives age-associated implantation failure in mouse embryos from aged females

doi: 10.1038/s41556-026-02052-1

Figure Lengend Snippet: a) Schematic of contractile inhibitor pathways to regulate potential trophectoderm spreading behaviors. We hypothesized that RhoA activator CN03 would phenocopy aged embryos, while ROCK inhibitor Ripasudil would rescue aged spreading behaviors. b) Representative images of Young (left) and Young-CN03 treated (right) embryos fixed and stained for actin. Violin plot shows a reduction of spread area in Young-CN03 treated embryos (Young N = 9, n = 63 implanting blastocysts; Young -CN03 N = 5, n = 25 implanting blastocysts). Scale bar is 50μm. Two tailed p values from Mann-Whitney test. c) Representative images of Aged (left) and Aged Ripasudil-treated (right) embryos fixed and stained for actin. Violin plot shows a rescue of spread area in Aged Ripasudil-treated embryos (Aged N = 4, n = 27 implanting blastocysts; Aged Ripasudil N = 3, n = 16 implanting blastocysts). Scale bar is 50μm. Two tailed p values from Mann-Whitney test.

Article Snippet: Embryos were treated with equilibrated KSOM-AA and/or IVC media containing 1μg/mL RhoA Activator II (Cytoskeleton Cat #CN03) or 10μM Ripasudil (Selleck Chemical Cat#50-136-6407) or 10 uM ResEcad (Sigma-Aldrich Cat#205615).

Techniques: Staining, Two Tailed Test, MANN-WHITNEY

Journal: Nature cell biology

Article Title: Elevated contractility drives age-associated implantation failure in mouse embryos from aged females

doi: 10.1038/s41556-026-02052-1

Figure Lengend Snippet: a) Schematic of trophectoderm spreading in which actomyosin generates contractile forces transmitted to other cells across E-cadherin–mediated junctions and to the substrate through integrin-based adhesions. b) Schematic of Trophectoderm Velocity Kymograph Analysis. TE contours were segmented at successive time points, and local displacements along the perimeter were used to compute edge velocities at each coordinate. These values were mapped over time to generate velocity kymographs, enabling spatial and temporal analyses of TE spreading. c) Embryos from aged mothers exhibit lower Trophectoderm Perimeter Velocity. Two tailed p values from Mann-Whitney Test. d) Perimeter outlines and Averaged trophectoderm velocity maps for Young (left) and Aged (right) conditions. Mural Trophectoderm perimeter outlines show the evolution of trophectoderm spreading over 24 hours. Perimeter outlines are color coded based on time post-implantation. Scale bar is 50 μm. Averaged TE Velocity Maps of normalized Trophectoderm contours for Young (n=20 implanting blastocysts) and Aged (n=15 implanting blastocysts). e) Schematic of pathway components contributing to tissue viscosity and reduced wetting behaviors. The cell-permeable drug CN03 activates RhoA for increased Myosin activity. Cell permeable ResEcad upregulates E-cadherin expression. Myosin-mediated forces at E-cadherin adhesions contribute to tissue viscosity and reduce tissue wetting. f) Quantification of Average Focal Adhesion Length per Embryo in Young (n=20 implanted embryos), Aged (n=17 implanted embryos), Young-CN03 treated (n=15 implanted embryos), and Young-ResEcad (n=12 implanted embryos) treated embryos (left). Two tailed p values from Mann-Whitney Test. Representative images of implanted embryos (right) demarcated with pMLC staining to show spread areas. Zoom images show localization of focal adhesion marker Paxillin at integrin adhesions. Scale bars are 50 μm and 5 μm, as indicated.

Article Snippet: Embryos were treated with equilibrated KSOM-AA and/or IVC media containing 1μg/mL RhoA Activator II (Cytoskeleton Cat #CN03) or 10μM Ripasudil (Selleck Chemical Cat#50-136-6407) or 10 uM ResEcad (Sigma-Aldrich Cat#205615).

Techniques: Two Tailed Test, MANN-WHITNEY, Viscosity, Activity Assay, Expressing, Staining, Marker