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. 2016 Jul 1;143(13):2376–2388. doi: 10.1242/dev.137984

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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. 4. — Sox2 deletion in IL progenitors results in reduction of POMC-positive cells, complete downregulation of PAX7, and ectopic differentiation of corticotrophs. (A) Immunofluorescence for SOX2, eYFP and POMC at 18.5 dpc in Nkx3.1^Cre mutants. POMC is present in SOX2^Low cells in the IL of Sox2^+/+;Nkx3.1^Cre/+;R26R^eYFP/+ embryos. In mutant Sox2^fl/fl;Nkx3.1^Cre/+;R26R^eYFP/+ embryos, SOX2 is essentially lost and the number of POMC-positive cells reduced. (B) Percentage of POMC-positive cells in the IL of Sox2^fl/fl;Nkx3.1^Cre/+ embryos at 18.5 dpc (23.2±6.2% of DAPI-positive cells, n=3), is significantly lower (***P=0.0008) than in heterozygous Sox2^fl/+;Nkx3.1^Cre/+ embryos (72.1±5.8%, n=3). Results presented as mean±s.d. (C) Immunofluorescence for PAX7 at 18.5 dpc. PAX7 is expressed exclusively in the IL in Sox2^fl/+;Nkx3.1^+/+ pituitaries. Expression is dramatically downregulated in Sox2^fl/fl;Nkx3.1^Cre/+ IL. (D) Immunofluorescence for POMC and glucocorticoid receptor (GR), at 18.5 dpc in Nkx3.1^Cre mutants. GR is normally present in POMC-positive corticotrophs in AL, but not in POMC-positive IL melanotrophs, as observed in Sox2^fl/+;Nkx3.1^Cre/+ pituitaries. In mutant Sox2^fl/fl;Nkx3.1^Cre/+ pituitaries, GR is ectopically present in the IL POMC-positive cells, demonstrating that these are in fact corticotrophs. Scale bars: 50 μm in A,C,D; 5 μm for magnifications in D. IL is outlined.