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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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(a) FACS cell-cycle analysis of hCD2⁺ cells from CTCF^fl/+ (n=4) and CTCF^fl/fl (n=5) mice after 48 h of stimulation with CD3/CD28 and T cells. Numbers indicate percentages ±s.d. Quantification of G1 and S/G2 phase proportions is shown on the right. p(G1)=0.0242; p(S/G2)=0.0221, two-tailed Student’s t-test. (b) Two-dimension expression plot. X axis, log2 fold values of significantly changed genes upon CTCF deficiency (CTCF^fl/fl versus CTCF^fl/+, CD3/CD28-stimulated B cells). Y axis, log2 fold values of significantly changed genes in PC versus GC B cells (data extracted from ref. 33. Coloured dots highlight genes of the PC differentiation programme. (c) Quantification of significantly changed cell-cycle-related genes. Log fold change (LogFC) in PC versus GC B cells (purple) and in CTCF^fl/fl versus CTCF^fl/+ CD3/CD28 T-stimulated B cells (orange). (d) RNA-seq analysis of CTCF expression in GC B cells and PC (data extracted from ref. 33). (e) qRT-PCR analysis of Blimp-1 (Prdm1) and Bcl-6 expression in CD3/CD28 T-stimulated CTCF^fl/+ (n=12) and CTCF^fl/fl (n=9) hCD2⁺ cells. P(Blimp-1)< 0.0001; P(Bcl-6)=0.0331. (f) CTCF binding at the Prdm1 locus in spleen B cells stimulated with LPS/IL4 (data from ref. 34). (g) CTCF ChIP-qPCR on Peaks A–C of the Prdm1 (Blimp-1) locus in spleen B cells stimulated with CD3/CD28 and T cells for 48 h (n=3). Data show per cent of input. IgG ChIP is shown as negative control. H19 Peaks 1–3 are shown as positive control for CTCF ChIP. Error bars indicate s.d. (h) RNA-seq analysis of Blimp-1 (Prmd1) targets in hCD2⁺ cells from CTCFfl/+ (n=3) and CTCFfl/fl (n=3) mice stimulated with CD3/CD28 and T cells for 48 h. (i) Blimp-1 binding at Myc, Cdkn1b, Rb1 and Ciita locus in plasmablasts (data from ref. 9) (j) ELISA quantification of IgM secretion from CD3/CD28 T-stimulated CTCF^fl/+ (n=3) and CTCF^fl/fl (n=3) cells for 72 h (P=0.0151). (CTCF^fl/+, white; CTCF^fl/fl; black). Statistical analysis was done with two-tailed unpaired Student’s t-test.