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. 2019 Sep 10;10:2143. doi: 10.3389/fimmu.2019.02143

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Copyright © 2019 Pellefigues, Mehta, Prout, Naidoo, Yumnam, Chandler, Chappell, Filbey, Camberis and Le Gros.

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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eYFP expression is robust during basophil degranulation and lifespan. (A,B) Whole splenocytes from Basoph8 mice were stimulated for 2 h with 100 ng/mL of PMA, ionomycin (Iono), or PMA + ionomycin (P/I), or left unstimulated (CT). CD49b+ CD200R3+ basophils expression of CD63 (A) and eYFP (B) is depicted in histograms and represented as mean fluorescence intensity (mfi) (n = 3). (C–G) 5,000 eYFP+ basophils were FACS sorted from the spleen of Hp infected mice at day 15, and stimulated with 1 ng/mL of IL-3, 100 ng/mL of ionomycin, or left unstimulated (n = 6). Their number (C) and phenotype was assessed post-sort, and at 1, 24, 48, and 72 h post stimulation. (D) The expression of eYFP by DAPI- eYFP+ basophils. (E) Representative contour plot showing the identification of eYFP+ living basophils left unstimulated at 24 h. (F) The expression of eYFP by DAPI- CD49b+ CD200R3+ basophils. (G) Representative contour plots showing the expression of eYFP by DAPI- CD49b+ CD200R3+ basophils in different conditions at different time points. Statistics are a one-way ANOVA with a Dunnett's comparison to the control value (A,B) Non-significant is not represented; ^*p < 0.05; ^****p < 0.0001. (D,F) Statistics are detailed in the text body for clarity. Data are representative of two independent experiments giving similar results (A,B) or show pooled data from two independent experiments (C–G).