Fig. 5. Effects of blocking SHH function on inner ear development. (A-D) Hybridoma cells expressing anti-SHH antibody (5E1) are pre-labeled with DiI, and implanted underneath the region of the hindbrain adjacent to the otocysts at E1.5. (A) A schematic diagram summarizing the procedure. (B) Bright-field and (C) fluorescent micrographs showing the location of implanted hybridoma cells (arrows) 24 hours after implantation. Asterisks in B,C,E,F indicate otocyst positions. (D) Ventral inner ear structures are missing in 5E1-implanted embryos (asterisk). (E-G) 5E1 cells are injected into the mesenchyme underneath the otocyst at E2.5 as indicated in E. (E) Bright-field and (F) fluorescent micrographs taken immediately after injection of 5E1 cells (arrows). (G) An inner ear injected with 5E1 at E2.5 lacks ventral structures (asterisk). Scale bars: in B, 500 µm for B,C; in E, 500 µm for E, F; in D, 500 µm for D,G.