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. 2021 Jun 23;11:13099. doi: 10.1038/s41598-021-92473-9

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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The renewal rate of circulating hemocytes. Crayfish were injected with one dose of EdU. HPT cells and circulating hemocytes were collected at different time points until 8 dpi. After fixation, cells were stained for EdU and analyzed by flow cytometry. Over 10,000 cells were analyzed for each sample. The percentages of EdU-incorporated cells in the HPT (A) and circulation (B) are shown with percentage-time scatter plots. Eight animals were analyzed at each time point. The relationships between time and EdU incorporation percentages were modeled using linear regression (1 ~ 3 dpi and 4 ~ 8 dpi for HPT cells; 3 ~ 6 dpi for circulating hemocytes). (C) Typical FL1-SSC scatterplots obtained in flow cytometry analysis (HPT cells, upper panel; hemocytes, lower panel). Cells in the right region of each plot are EdU-incorporated.