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. 2018 Dec 5;8:17651. doi: 10.1038/s41598-018-36121-9

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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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Signaling downstream of the BCR is mostly unaffected by cryopreservation. B cells were isolated from healthy blood donors and analyzed immediately (n = 3; fresh) or after cryopreservation (n = 3; cryo). The cells were stimulated with anti-IgM (10 μg/mL) for the indicated time-course before fixation and phospho flow analysis as described in Methods. Results are shown as mean ± standard error of the mean (SEM). Statistical significance was calculated between the two groups (fresh, cryo) by a paired t-test with excluded time parameter. Significant p-values are indicated.