Impaired lymphatic valve protrusion in Tie2Cre;Piezo1cKO mice.

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Impaired lymphatic valve protrusion in Tie2Cre;Piezo1cKO mice. Impaired lymphatic valve protrusion in Tie2Cre;Piezo1cKO mice. (A) E18.5 mesentery with Prox1 staining. (Scale bar: 100 μm.) (B) Number of VFRs (containing n ≥ 10 Prox1high nuclei) per area in E18.5 mesentery. Mean ± SEM (n = 4 for WT, n = 5 for Tie2Cre;Piezo1cKO embryos). P = 0.35, unpaired t test. (C) Stages of lymphatic valve formation. Stage 2: there are n < 5 reoriented Prox1high nuclei and the vessel is not constricted. Stage 2.5: n ≥ 5 Prox1high nuclei are reoriented but the vessel is not constricted. Stages 3 and 4: n ≥ 5 Prox1high nuclei are reoriented and the vessel is constricted. (D) Percentage of each stage among VFRs in E18.5 mesentery. n = 17 VFRs from four embryos per genotype. *P < 0.05, χ2 test. (E) Representative images of VFRs in E18.5 mesentery. Yellow lines (Top, side view) show where images were optically cross-sectioned (Middle) and schematic illustration of distribution of Prox1high nucleus (green), VE-cadherin (red), and F-actin (blue) signals (Bottom). Dotted lines demarcate lymphatic vessels. (Scale bar: 100 μm.) (F–H) Percentage of VFRs in E18.5 mesentery in which several Prox1high nuclei (F), VE-cadherin (G), or F-actin (H) signal was located in the more central regions than Prox1high nuclei on the vessel wall. Right shows examples of patterns of each signal. n = 10 images from four WT embryos, n = 14 images from four Tie2Cre;Piezo1cKO embryos. *P < 0.05, χ2 test. Keiko Nonomura et al. PNAS doi:10.1073/pnas.1817070115 ©2018 by National Academy of Sciences