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. 2020 Sep 9;11:531785. doi: 10.3389/fimmu.2020.531785

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Copyright © 2020 Zhou, Xing, Tang, Sheng, Chi and Zhan.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Co-localization analysis of EdU and CD4 in the spleen of flounder. (A) Indirect immunofluorescence of EdU and CD4 in flounder spleen after PHA + rHis, PHA + rHis-IL-2 injection on day 7; bar = 100μm. The unimmunized serum was used as primary antibody for negative control. (B) Image analysis for co-localization of EdU and CD4 in a selected area of spleen using Image J software. (a) A new merged image showing the PHA + rHis-IL-2 group in Figure 4A. Compared with the above image, magenta was represented CD4-positive signals, aqua was represented EdU-positive signals, blue was represented EdU and CD4 double-positive signals, respectively. (b) The interactive 3D surface plot of the selected area (the position of the rectangle) of (a). (c) Analysis of co-localization of EdU and CD4 according to the optical density of three fluorescence in the selected area (the position of the rectangle of a). 15 fish per group.