DEV000760 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Expression patterns of Ptc1 and Gli1, Gli2 and Gli3 transcription factors in the developing inner ears at 9.5 and 12.5 dpc. Expression patterns of Ptc1 (A,B), Gli1 (C,D), Gli2 (E,F) and Gli3 (G,H) in the developing inner ears at 9.5 (A,C,E,G) and 12.5 (B,D,F,H) dpc. Graded expression patterns of Ptc1 (A,B) and Gli1 (C,D) in the inner ear are evident at 9.5 dpc and these patterns are maintained at 12.5 dpc. Gli2 transcripts are detected throughout the inner ear, except the dorsal-medial area of the otocyst at 9.5 and the endolymphatic duct area at 12.5 dpc (E,F). Gli3 is broadly expressed in the inner ear and periotic mesenchyme at 9.5 and 12.5 dpc (G,H). vp, vertical pouch; co, cochlea; hp, horizontal pouch, D, dorsal; L, lateral. Scale bars: in A, 100 μm for A,C,E,G; in B, 100 μm for B, D, F, H.

• Supplemental Figure 2 -

  Fig. S2. Expression patterns of Gli2 and Gli3 in the otocysts of Shh^−/−;Gli3^+/− and Shh^−/−;Gli3^−/− embryos. Gli2 (A,B,C) and Gli3 (D,E) expression patterns in Shh^+/−;Gli3^+/− (A,D), Shh^−/−;Gli3^+/− (B,E) and Shh^−/−;Gli3^−/− (C) inner ears. Gli2 transcripts are present in most of the inner ear, but only weakly detectable in the endolymphatic duct (ed, arrow) region of all three mouse strains (A,B,C). The broad Gli3 expression in the inner ear of Shh^−/−;Gli3^+/− (E) is also similar to that in Shh^+/−;Gli3^+/− embryos (D). Orientations: D, dorsal; L, lateral. Scale bar: 100 μm

• Supplemental Figure 3 -

  Fig. S3. Shortened cochlear duct in Shh^−/−;Gli3Δ699/− mutant inner ears. Three representative regions from serial sections of Shh^+/−;Gli3Δ699/− (A,B,C) and Shh^−/−;Gli3Δ699/− (A′,B′,C′) inner ears at 12.5 dpc (see schematic). At the canal level (A,A′) the morphologies are similar between both ears. At the saccular/proximal cochlear duct level (B,B′), a rudimentary saccule is present in Shh^−/−;Gli3Δ699/− inner ear, similar to those in Shh^+/−;Gli3Δ699/− ears. However, below the level of the saccule, no cochlear tissue resembling those in Shh^+/−;Gli3Δ699/− (E) is present in Shh^−/−;Gli3Δ699/− inner ears (F). Overall, the inner ear morphology of Shh^−/−;Gli3Δ699/− mutants is similar to Shh^−/−;Gli3^+/− ears (see Fig. 3C). asc, anterior semicircular canal; cd, cochlear duct; ed, endolymphatic duct; lsc, lateral semicircular canal; psc, posterior semicircular canal; sac, saccule, A, anterior; L, lateral. Scale bar: 100 μm