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. 2020 Mar 18;9:e52779. doi: 10.7554/eLife.52779

Figure 1. Characterization of Piezo1 and Piezo2 expression in the developing long bones.

(a) Expression of Piezo1 and Piezo2 examined by whole-mount in situ hybridization of embryonic limb buds at the indicated stages. (b) Immunostaining for tdTomato, Sp7 and Sox9 in consecutive humerus sections of the Piezo1-tdTomato forelimb buds. Arrows: Membranous tdTomato and nuclear Sp7 were found in the differentiating osteoblasts and hypertrophic chondrocytes in the same regions. Arrowheads: Extra-skeletal Piezo1-tdTomato expression. (c) Representative image of GFP and tdTomato fluorescent staining of humerus sections from the Piezo1-tdTomato; Sp7-Cre::GFP mouse at P0. The boxed region was shown in higher magnification in the right panel. Arrows: colocalization of tdTomato and GFP in the differentiating osteoblasts. Arrowheads: Hypertrophic chondrocyte or extra-skeletal expression of Piezo1-tdTomato. (d) Representative images of tdTomato costained with Sp7 or Sox9 on consecutive humerus sections from the E13.5 Piezo2Cre; Rosa26-tdTomato limb bud. Arrows: TdTomato costained with Sp7 in the differentiating osteoblasts. Arrowheads: Joint or extra-skeletal Piezo1-tdTomato expression. All scale bars, 100 μm. DAPI (blue) stain the nucleus.

Figure 1.

Figure 1—figure supplement 1. Mouse Piezo1 and Piezo2 expression.

Figure 1—figure supplement 1.

(a, b) Expression of Piezo1 and Piezo2 examined by whole mount in situ hybridization of mouse embryos at E10.5 and E12.5 with Piezo1 and Piezo2 sense or antisense probes. (b) Ventral view of Piezo2 expression in the limb bud at the indicated stages. (c) Section in situ hybridization by RNAscope to detect Piezo1 mRNA expression in E16.5 humerus. Arrows: Piezo1 expression in the differentiating osteoblasts and muscle. Arrowheads: stronger expression of Piezo1 in the non-muscle connective tissues. Boxed regions are shown in higher magnification on the right panel. Scale bar, 100 μm. (d and e) Fluorescent images of tdTomato with costained Sp7 in humerus of the Piezo2Cre; Rosa26-tdTomato mouse at P14 (d) and E17.5 (e). Boxed region in (e) is shown on the right side. Some of the Piezo 2 lineage cells expressed Sp7. Scale bars, 100 μm. DAPI stains the nucleus.