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. 2016 Feb 15;5(3):279–288. doi: 10.1242/bio.015032

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© 2016. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 5. — In the absence of En1 the IsO is disorganized. (A-D,I-L) Otx2 expression is present in midbrain at E12.5, but is caudally extended into the hindbrain in En1KO and in En1CreKO, in a mosaic fashion (arrowheads). (E-H,V-Y) Wnt1 expression is present in midbrain at E12.5, but is caudally extended into the hindbrain in En1KO, in a mosaic fashion (arrowheads). (i-ii) A pseudo-overlay reveals that the ectopic, mosaic expression of Otx2 (green) and Wnt1 (red) in En1KO is mutually overlapping. (M-P) Th expression is found ectopically in the YFP-positive R1 at E12.5 in En1CreKO in (para)medial sections. (iii-iv) The pseudo-overlay of Otx2 (green) and Th (red) displays caudal ectopic expression in the En1CreKO. (R-U) Fgf8 expression is present at the IsO in wild-type at E12.5, but is lost in medial sections in the En1KO and caudally multiplied in lateral sections (arrowheads). (v-vi) The pseudo-overlay of Fgf8 (green) and Wnt1 (red) displays caudal ectopic expression in the En1KO in a mutually exclusive manner. (Z-GG) β-galactosidase activity is present in IsO at E14.5 in En1WT;BatGal+ animals, and is caudally extended in the En1KO;BatGal+ (arrowheads in BB,CC). Adjacent sections of Wnt1 mirror the β-galactosidase activity, both the restricted expression at the IsO in control and the enlarged area in En1KO;BatGal+ animals (arrowheads in FF). (vii-viii) The pseudo-overlay of β-galactosidase activity (green) and Wnt1 (red) displays caudal and overlapping ectopic expression in the En1KO;BatGal+. Scale bar: 100 μm.