DEV01259 Supplemental Figures

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Anti-SOX9 antibodies are specific for the chick SOX9 protein. (A) Western-blot analyses using anti-SOX9 (top panel) and anti-tubulin (bottom panel) antibodies. Cell lysates were prepared from human NT2/D1 (N-Tera2, clone D1, a human pluripotent embryonic carcinoma cell line) (1), chick embryo harvested at stage 16 (2), guts dissected from 5-day-old embryos (3). Anti-SOX9 antibodies detect a single band corresponding to the chick SOX9 protein both in embryo and gut lysates, which migrates with a lower calculated molecular weight compared with the human SOX9 protein as previously reported (Kamachi et al., 1999). (B-J) Immunohistochemical analyses on chick stage 26 paraffin cross-sections using anti-SOX9 antibodies. Specific detection of nuclear SOX9 staining into the prevertebrate structure (B,H) and the neural tube (E). (C,F,I) Addition of GST peptide antigen (a-SOX9 + GST peptide) does not affect anti-SOX9 immunoreactivity. (D,G,J) Addition of SOX9 transactivation domain peptide antigen in fusion with GST (a-SOX9 + SOX9 peptide) at the same concentration eliminates anti-SOX9 immunoreactivity.

• Supplemental Figure 2 -

  Fig. S2. Characterization of RCAS-SOX9 constructs. (A-C) Immunohistochemistry detection using anti-SOX9 antibodies on RCAS-GFP (A), RCAS-SOX9 (B) transfected CEF cells and RCAS-SOX9 infected E7 stomach (C). SOX9 protein is expressed in RCAS-SOX9-transfected CEF cells (red arrow, B) and there is ectopic SOX9 expression in the mesenchyme of the gizzard (red arrow, C). CEF, chick embryonic fibroblast; e, endoderm; gizz, gizzard; m, mesoderm.

  Reference

  Kamachi, Y., Cheah, K. S. and Kondoh, H. (1999). Mechanism of regulatory target selection by the SOX high-mobility-group domain proteins as revealed by comparison of SOX1/2/3 and SOX9. Mol. Cell. Biol. 19, 107-120.