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. 2018 Sep 12;9:3691. doi: 10.1038/s41467-018-06033-3

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© The Author(s) 2018

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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Fig. 6 — SG neuron types in new born mice and comparative analysis of their transcriptome with adult SG neurons. a t-SNE of SG neurons showing four different clusters at P3. b Violin plots showing the expression of marker genes in log-transformed scale. c In vivo validation of the identified neuron types by immunohistochemical and fluorescent in situ hybridization using identified marker genes in P3 cochlea. Type II SG neurons were identified by Peripherin (Peri), Plk5, Etv4 (Etv4^GFP transgenic mouse), TH, and Gabrg2. Ia neurons were identified by Pou4f1, Runx1, and CR. Ib neurons were identified by Runx1 and Pou4f1 and Ic neurons, by Rxrg, Pcdh20, and CR expression. Lypd1 and Calb1 expression could not distinguish type Ia from Ib neurons at this stage. Note that co-localization on sections could never be observed for markers expressed in different populations of neurons in the scNRAseq data. d Gene set enrichment analysis of P3 type I and type II SG neurons visualized by network. e Correlation analysis of SG neuron types from adult and P3 stages, using average expression of all differentially expressed genes as input. f, g Visualization of SG neuron types from adult and P3 stages using tSNE, revealing the conserved subclass identities between the two samples. h Volcano plots of gene expression differences between adult and P3 SG neuron types for type Ia (top panel) and type II (bottom panel). Genes differentially expressed in adult or P3 are marked by red or blue dots respectively. Scale bar: 20 μm (c)