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. 2019 Apr 16;9:6118. doi: 10.1038/s41598-019-42630-y

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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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GSCs are competed out of the niche by CySCs and differentiated in c587^ts > tkv^CA testis. Time chase experiment is carried out to trace the fate of GSCs. Time points examined are indicated. The hub is marked by white asterisk, GSCs are indicated by yellow arrowheads, and CySCs by white arrowheads. CySCs begin to closely attach to the hub on the 2^nd day at 29 °C from 18 °C. The hub is closely associated by CySCs on the 3^rd day after shifting, and all GSCs are competed out of the niche by CySCs and undergo differentiation (white arrowhead). Germline cells move further away from the hub and undergo differentiation by the 6^th day. Almost all the spermatogonia are terminally differentiated, and fully differentiated spermatids can be observed (yellow arrowhead). GFP in green, blue indicates DAPI staining for DNA. Scale bars: 10 μm.