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. 2020 Nov 12;10:19696. doi: 10.1038/s41598-020-76677-z

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© The Author(s) 2020

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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Entry of hemocytes in mussels. CFSE-stained hemocytes from Mytilus spp. were added to seawater. After the indicated times, IVF and hemolymph were collected and analyzed for the presence of CFSE-positive cells. (A) A representative contour plot showing hemocytes collected from IVF of mussels incubated without (left) and with CFSE-positive hemocytes (right). In the upper corner of the left contour plot, a representative overlay histogram of unstained (control) hemocytes in blue and CFSE stained hemocytes in green, (B) Effect of temperature on entry of CFSE-positive cells in IVF of mussels. (C) Percentage of CFSE-positive cells recovered in IVF of mussels at different temperatures 90 min after addition of CFSE-stained cells. The autofluorescence controls are shown in red. Its shows the autofluorescence as measured in (A) generated by unstained hemocytes recovered in IVF of mussels incubated in the same conditions. (D) Percentages of CFSE-positive cells present in hemolymph of Mytilus spp. maintained at 23 °C for 90 min after addition of CFSE-positive cells. The dotted line represents the average of CFSE-positive cells in hemolymph. The data are representative of two independent experiments.