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. 2020 Aug;109:103715. doi: 10.1016/j.dci.2020.103715

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© 2020 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 4 — Percentage of large and small IgM⁺ B cells in (A) spleen and (B) kidney 15 and 30 days after the anal administration of antigens. Leukocytes isolated from spleen and kidney of rainbow trout stimulated anally with the different antigens were incubated with an anti-trout IgM mAb conjugated to phycoerythrin (PE) and analyzed by flow cytometry. Cells were gated as lymphoid on the basis of their FSC and SSC and percentages of small (naïve) and large (activated) IgM⁺ B cells were determined on singlet, live (DAPI negative) IgM⁺ B cells. Representative dot plots showing the percentage of small and large IgM⁺ B cells are included for each group and tissue at both sampling times. Graphs on the right show the percentage of large IgM⁺ B cells (mean +SD, n = 7). Asterisks denote significantly different values between TNP stimulated groups compared with control group (*p ≤ 0.05 and ***p ≤ 0.001).