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Fig. 2. Mash1 expressing progenitors give rise to dILA and dILB neurons. (A) Strategy used to replace the Mash1-coding sequence by GFP. The wild type Mash1 locus, the targeting vector, and the recombined allele before (Mash1^GFPneo) and after deletion of the neo resistance cassette (Mash1^GFP) are depicted. In the targeting vector, the GFP sequence (green) was fused to the initiation codon of Mash1, and the coding sequence of Mash1 (blue) was deleted. No additional polyA was introduced, and the exon-intron structure of the Mash1^GFP allele is identical to the one of wild-type Mash1. (B,C) In situ hybridization of consecutive sections from a E12.5 Mash1^GFP/+ spinal cord using probes directed against Mash1-coding sequence (B) or GFP (C). The expression patterns are identical. (D-F) Immunohistochemical analysis of spinal cords of Mash1^GFP/+ mice at E12.5 with antibodies directed against (D) Lbx1 (red), GFP (green) and Mash1 (blue), (E) Lmx1b (red) and GFP (green), and (F) Lhx1/5 (red) and GFP (green). The spinal cord is shown at low (D) and high (E,F) magnification. All Lmx1b+ (dILB) neurons are also GFP+ in E; all Lhx1/5 (dILA) neurons are also GFP+ in F. Scale bars: 100 µm in B-D; 10 µm in E,F.

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