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. 2017 Jul 19;7:5922. doi: 10.1038/s41598-017-06397-4

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© The Author(s) 2017

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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CD300f negatively regulates eosinophil-derived IL-4 production and governs IL-5-induced ERK and pAKT phosphorylation. The mRNA (A) expression of IL-4 was assessed in the white and brown adipose tissue (WAT and BAT, respectively) by quantitative PCR and normalized to the house keeping gene hypoxanthine-guanine phosphoribosyltransferase (Hprt). Protein expression of IL-4 (B) was determined by ELISA. In (C), primary eosinophils were purified from the WAT of Il5 ^Tg and Il5 ^Tg /Cd300f ^−/− mice and either left untreated (NT) or activated with phorbol 12- myristate 13-acetate (PMA). Thereafter, secretion of IL-4 was determined in the culture supernatants (C). The expression of CD300f (D) and IL-5 receptor α (IL-5Rα) (G) in I.29 B cells is shown following viral infection with empty vector or CD300f-containing vector (D). Following retroviral infection with empty vector of CD300f-containg vector, I.29 cells were stimulated with IL-5 for the indicated time points (E,F) and the phosphorylation of ERK (p42/44) (E,F) and AKT (H) was assessed by phosphoflow (E,F) and western blot, respectively (H). Data in (A,B) are from n = 5 mice, in (C) from n = 3 independent experiments, in (D,H) from n = 4 independent experiments; *p < 0.05, **p < 0.01, ***p < 0.001 as analyzed by Student’s t-test (A–C) and two-way Anova followed by Tukey post-hoc test (F).