Figure 4
From: Engineered three-dimensional rabbit oral epithelial–mesenchymal–muscular hybrid sheets
Alteration of expressed basement membrane components in hybrid sheets. (a) Double staining of rabbit hybrid sheets after 1, 3, 5, and 7 days with laminin (green) and desmin (red). Nuclei were stained with DAPI. Scale bars, 50 μm. (b) Double staining of rabbit hybrid sheets after 1, 3, 5, and 7 days with collagen type IV (Coll IV, green) and desmin (red). Nuclei were stained with DAPI. Scale bars, 50 μm. (c) Coll IV western blot of rabbit hybrid sheets after 1, 3, 5, and 7 days. β-Actin was used as an internal control. (d) Relative expression of Coll IV (Coll IV/β-actin) protein (n=4). (e) Laser scanning confocal images of a hybrid sheet after 7 days. All panels are 1 μm scanning images. Coll IV (green) was expressed in the epithelial basement membrane, the collagen gel layer containing rOMMCs, and in the desmin (red)-positive rOMYCs layer in all images. Nuclei were stained with DAPI. Scale bars, 20 μm. (f) Representative image of e. Coll IV was expressed in fused and multi-nucleated rOMYCs (arrowheads). Light upper insert is an enlarged image of the area depicted by arrowheads. DAPI, 4,6-diamidino-2-phenylindole; rOMYCs, rabbit oral myoblasts.
