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. 2016 Sep 1;129(17):3295–3308. doi: 10.1242/jcs.186015

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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. 2. — PPFs are correctly targeted by phrenic nerves. Whole-mount immunohistochemistry against eGFP-tagged Hb9 (somatic motor projections, green) and neurofilaments (motor and sensory axons, red) at stage E11.5 (A–C) and E12.5 (A′–C′). Control embryos show fasciculated phrenic projections after the brachial plexus at E11.5 (A, arrowhead). At the same stage, Npn1^Sema−/− (B) and Npn-1^cond−/−;Olig2-Cre⁺ mutant embryos (C) expose severely disorganized projections within the brachial plexus region (asterisks). Phrenic axons reach the developing diaphragm as a fasciculated nerve branch regardless of the genotype (A–C, arrowheads). The phrenic nerve correctly targets the primordial developing diaphragm and starts to branch dorsally and ventrally at E12.5 (A′–C′, arrows). H, heart; L, liver; Lu, lung. Scale bars: 200 µm.