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Fig. 5. In reaggregated explants cells expressing pre-ZLI genes exhibit Shh-dependent segregation behaviors: (a) Experimental design: AAC were prepared from embryos injected as indicated. At st. 8/9 ßgal and gfp cells are dissociated and reaggregated to generate explants made of Gfp/ßGal (red) mixed cell types. The explants are cultured for 48 h. (b, c) Representative explants composed of mixed cells expressing the different neuroepithelial genes as indicated are shown (b) without N-Shh (c) with N-Shh (100%, n≥10 for each condition). The % of explants showing the phenotype is indicated. The scale bar stands for 0.5 mm. (A) AAC cells do not segregate from Otx2-expressing cells. AACs cells expressing otx2 spread randomly when mixed with AACs cells (red) (b) in the absence (100%, n=12) and (c) in the presence of N-Shh (100%, n=10). (B) Pre-ZLI-like cells segregate from Otx2-expressing cells. Pre-ZLI cells (red) regrouped and separated from anteriorized neuroepithelial cells in the absence (b) and presence (c) of N-Shh (100%, n=10 for each condition). (F) In the presence of N-Shh pre-ZLI-like cells segregate from thalamus-like cells (b, c) pre-ZLI cells (red) regrouped, and separated from thalamus-like cells in the presence (c) but not in the absence (b) of N-Shh (100%, n=10 for each condition). Image published in: Juraver-Geslin HA et al. (2014) Copyright © 2014. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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