DEV025304 Supplementary Material

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• Supplemental Figure S1 -

  Fig. S1. Expression of Osr2, Pax9, Fgfr2, Tgfb3, Shh and Patch1 in the developing palatal shelves is not altered in the Mn1^−/− mutant embryos. (A-C) The expression of Osr2 (A), Pax9 (B) and Fgfr2 (C) in wild-type embryos at E13.5. Osr2 and Pax9 are strongly expressed in the palatal mesenchyme, whereas Fgfr2 is expressed in the palatal epithelium and medial palatal mesenchyme. (D-F) The expression of Osr2 (D), Pax9 (E) and Fgfr2 (F) in Mn1^-/- mutant littermates at E13.5. The expression of Osr2, Pax9 and Fgfr2 is not altered in Mn1^−/− mutant embryos. (G-I) The expression of Tgfb3 (G), Shh (H) and Patch1 (I) in wild-type embryos at E13.5. The expression of Tgfb3 and Shh is restricted in the palatal epithelium, whereas Patch1 is expressed in the palatal mesenchyme. (J-L) The expression of Tgfb3 (J), Shh (K) and Patch1 (L) in Mn1^−/− mutant embryos is not altered in Mn1^−/− mutant embryos in comparison with the wild-type littermates. p, palatal shelf; t, tongue.

• Supplemental Figure S2 -

  Fig. S2. Schematic representation of the genomic organization corresponding to alternative transcripts from the human TBX22 and mouse Tbx22 genes, respectively. (A) The human TBX22 gene contains two alternative promoters, hP0 and hP1, resulting in two TBX22 transcripts differing the 5′ untranslated reigon. The predominant TBX22 transcript in human embryos is that containing Eexons 0-8, transcribed from the distal hP0 promoter, approximately 10 kb upstream from exon 1. The other transcript is expressed at a very low level, and the putative proximal hP1 promoter was not active in an in vitro promoter assay (Andreou et al., 2007). Exons are boxed and numbered, whereas the promoter fragments used for in vitro assays are marked in oval shape. (B) 5′RACE experiments revealed that the mouse embryos express three distinct Tbx22 transcripts that differ only in the 5′ untranslated sequences. Exons 1a or 1b can be part of a transcript under the control of the proximal promoter mP1. Exon 0, which lies ∼8 kb upstream of exon 1, splices onto a splice site inside exon 1 to express a transcript under the control of the distal promoter mP0.

• Supplemental Figure S3 -

  Fig. S3. Sequences for mouse Tbx22 mP0, mP1 and human TBX22 hP0, hP1 promoter regions. Lower case letters represent non-coding upstream genomic sequences, whereas upper case is exonic sequence. Primer sequences used to amplify promoter fragments for the promoter-luciferase constructs are underlined. Start codon in exon 1 (ATG) is boxed. Each putative Mn1 binding core sequence (CACCC or GGGTG) is highlighted in yellow. The mP0 and mP1 sequences match exactly the mouse Tbx22 genomic sequences in the NCBI Mouse Genome Database, whereas the hP0 and hP1 sequences match corresponding human TBX22 genomic sequences exactly the NCBI Human Genome Datase. Each sequence has also been deposited into GenBank.