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. 2019 Mar 20;9:4936. doi: 10.1038/s41598-019-39996-4

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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C boutons on brainstem motor neurons are Pitx2 derived. In green anti-ChAT antibody labels motor neuron somata and cholinergic C boutons on them, in red anti-dsRed antibody labels tdTomato+ Pitx2 derived synapses. (A–D) The first column contains images of the cranial motor nuclei and the next three columns contain close up images of motor neurons (merge, red only, green only). (Ai–iv) Pitx2 derived (red) C boutons (green) on the facial nucleus (green) appear yellow. (Bi–iv) Nucleus abducens does not receive C bouton inputs. (Ci–iv) Pitx2 derived (red) C boutons (green) on the trigeminal nucleus (green) appear yellow. (Di–iv) Trochlear and oculomotor nuclei do not receive C bouton inputs. Scale bar (A–D) 10 μm. (Ei) Half section of medulla showcasing the position of facial nucleus, (Eii) nucleus abducens, (Eiii) trigeminal nucleus, (Eiv) trochlear/oculomotor nucleus, in white boxes. Scale bar (E) 100 μm.