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. 2018 Feb 26;8:3580. doi: 10.1038/s41598-018-22044-y

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© The Author(s) 2018

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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Phenotypic analysis of CD4⁺ T cells after immunization by MntC. (A) After 7 days’ incubation, lymphocytes were collected and treated with PMA, ionomycin and Golgistop for 6 hours. Then these cells were washed and stained with fluorescent antibody for Th1, Th17 and Th2, analyzing by flow cytometry. (B) Th1, Th17 and Th2 cell responses of all immunized mice were statistically analysed. The grey bars represent the proportion of double-positive cells. Significant differences were indicated by (*P < 0.05), (**P < 0.01) and (***P < 0.001).