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. 2017 Jul 5;8:16067. doi: 10.1038/ncomms16067

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Copyright © 2017, The Author(s)

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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CTCF^fl/+ (n=4) and CTCF^fl/fl (n=5) splenic B cells stimulated with CD3/CD28 T cells were treated with anti-CD40 antibody (+anti-CD40), and compared to cells left untreated (−anti-CD40). (a) Representation of the experimental approach. (b) Analysis of Blimp-1 (Prdm1) mRNA by qRT-PCR in hCD2⁺ CTCF^fl/+ or CTCF^fl/fl cells after 48 h without or with anti-CD40 treatment. ***P=0.0001, *P=0.0159. (c) Histograms show the percentage of B220⁺ B cells in CTCF^fl/+ and CTCF^fl/fl B-T CD3/CD28 co-cultures after 48 h of anti-CD40 treatment. Lower bar graph shows quantification of B220 proportion at 24 and 48 h after CD40 treatment. **P=0.0063, *P=0.023. (d) Histograms show the percentage of cycling (S+G2/M) cells in CTCF^fl/+ and CTCF^fl/fl B–T CD3/CD28 co-cultures after 48 h of anti-CD40 treatment. Lower bar graph shows quantification of cycling cells at 24 and 48 h after CD40 treatment. **P=0.0083, *P=0.0126. Mean values±s.d. are shown. Statistical analysis was done with two-tailed unpaired Student’s t-test.