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. 2019 Dec 4;10:5530. doi: 10.1038/s41467-019-13157-7

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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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Fig. 3 — Study of the origin of dividing HCs and SCs in adult cochlea by lineage tracing. a A schematic diagram depicting the in vivo experimental procedure. b In control adult Sox2-CreER/tdT^f/f mice 10 days after ad-V5 injection and Tamoxifen (Tm) treatment, all IHCs were tdT negative and were surrounded by tdT⁺ SCs. No EdU labeling was detected. c Following Tm treatment and ad-Myc/ad-NICD injection into adult Sox2-CreER/tdT^f/f mouse cochlea for 10 days, dividing IHCs were detected (arrows). All IHCs (dividing, arrows; non-dividing, arrowhead) were tdT negative. d Schematic diagram of lineage-tracing of proliferating adult SCs in vitro. e–g EdU labeling in the SCs (tdT⁺) in cultured adult Sox2-CreER/tdT^f/f cochlea after treatment with Tm and infection with ad-Myc/ad-NICD. Judging by their locations, likely dividing Deiters’ cells (arrow) and pillar cells (arrowhead) were identified. Bracket indicates the sensory epithelium (SE). h–j MKI67 labeling in the SCs (tdT⁺) in cultured adult Sox2-CreER/tdT^f/f cochlea after treatment with Tm and infection with ad-Myc/ad-NICD. k A confocal image of the surface view of the middle turn of adult Sox2-CreER/tdT^f/f cochlea after treatment with Tm and infection by ad-Myc/ad-NICD to show numerous proliferating SCs (tdT⁺/EdU⁺) in the SE region. l, m z-axis cross-section views of confocal images in (k) showed dividing SCs by triple labeling of tdT⁺/EdU⁺/SOX2⁺ (likely Deiters’ cells, arrows; Hensen cell, arrowhead. tdT: red; EdU: green; SOX2: white). Dashed lines mark the basilar membrane and the tunnel of Corti (TC). Scale bars: 50 μm in (k); 10 μm in other figures.