DEV005868 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Characterization of Foxn4 expression in the embryonic chick spinal cord. All images are of single or double in situ hybridization signals. The number of Foxn4-positive cells in the ventral cord increases between st13 and st22 (A-C). At all stages, the Foxn4-expressing cells are in the VZ or at the lateral border of the VZ (e.g. C). (D) Nkx6.1 (red)-Foxn4 (green) at st22. Foxn4-positive cells all fall within the Nkx6.1 expression domain, which marks the dorsal limit of the p2 progenitor domain. (E) Gata2 (red)-Foxn4 (green) at st25. Many cells are double-positive. A higher magnification image of the indicated area in E is reproduced in G, showing that Gata2 persists longer in the V2b lineage than Foxn4, so that there are more Gata2-only cells in more lateral positions, further from the lumen. In G, the boxed cells at the ventricular surface are reproduced, fluorescence channels separated, at the top right of the panel. (F) Chx10 (red)-Foxn4 (green) at st22. A higher-magnification image of the indicated area in F is reproduced in H, showing that there is no co-localization between Chx10 and Foxn4.

• Supplemental Figure S2 -

  Fig. S2. Ectopic Foxn4 expression represses interneuron fates other than V2a. Chick embryos were electroporated at st14 with β-actin-Foxn4-IRES-GFP and analyzed after 24 hours by double immunolabelling with anti-GFP (green) and either anti-En1 or anti-Lhx1/5 (‘anti-Lim1+2’; Developmental Studies Hybridoma Bank; red). The numbers of both En1-positive (A) and Lhx1/5-positive (B) cells were reduced (see text for quantification).