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

FIGURE 4.

FIGURE 4

Co-localization analysis of EdU and CD4 in the gill of flounder. (A) Indirect immunofluorescence of EdU and CD4 in flounder gill after PHA + rHis, PHA + rHis-IL-2 injection at day 7; bar = 200μm. The cell nuclei were counterstained in blue with DAPI. 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 gill using Image J software. (a) A new merged image showing the PHA + rHis-IL-2 group in (A). 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 panel (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)]. Fifteen fish per group.