Fig. 7

Fig. 7 Hhip regulates Smoothened localization. (A–D) Smo is detected intracellularly (B, asterisk) and at the cell surface (B, arrow and broken line) in cells near the notochord (A, B, D, 93%, n = 16) compared with lateral cells that contain primarily intracellular Smo (C, 50%, n = 6). (E, F, G, H) hhip-MO suppresses internalization of Smo in medial cells (83%, n = 18). (I, J, K, L) Inhibition of endocytosis suppresses internalization of Smo (100%, n = 7). 100 μM MDC added at the 40% epiboly stage and then incubated to the 8-somite stage. (A–L) Embryos were injected with myc-tagged smo plasmid at the 1-cell stage and incubated to the 8-somite stage. Single confocal images of myc-tagged Smo (A–G, I, J, K and D, H, L, upper panels). Embryos were labeled with anti-myc antibody and myod (A, E, I) or anti-β-catenin antibody (D, H, L), a marker of the membrane (Nagafuchi, 2001). (D, H, L, Lower panels) Confocal micrographs (transverse z series) through myc-tagged Smo expressing cells. (A, E, I) Arrows indicate the cells shown at higher magnification in right panels. (B, C, D, F, G, H, J, K) Asterisks and arrows indicate intracellular and surface localization of Smo, respectively. N, notochord, scale bar: (A, E, I) 10 μm, (B, C, D, F, G, H, J, K, L) 2.5 μm.