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. 2020 Nov 18;11:5894. doi: 10.1038/s41467-020-19692-y

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© The Author(s) 2020

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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Fig. 6 — Nuclear and cytoplasmic subcellular fractions were obtained from Treg and Th17 cells and tested for antisense expression. a, b Mean ± SEM CD39-AS RNA levels in Treg and Th17 cells from HS (n = 7) and Crohn’s disease patients (n = 12) (a **P = 0.008 for HS and *P = 0.01 for Crohn’s patients; b *P = 0.01 for HS and **P = 0.009 for Crohn’s patients using two-sided unpaired t test). Based on the results of mass spectrometry, Treg and Th17 cells, differentiated from the peripheral blood of healthy subjects and Crohn’s disease patients, were exposed to scramble or gene-specific siRNA. c, d Mean ± SEM CD39 and CD39-AS RNA levels and CD39 MFI in untreated and siRNA (or scramble) treated Treg and Th17 cells from healthy subjects (HS; Treg: CD39 mRNA n = 6; CD39-AS RNA n = 5; CD39 MFI n = 10; Th17: CD39 mRNA n = 6; CD39-AS RNA n = 7; CD39 MFI n = 10) and Crohn’s disease patients (Treg: CD39 mRNA n = 6; CD39-AS RNA n = 4; CD39 MFI n = 6; Th17: CD39 mRNA n = 5; CD39-AS RNA n = 4; CD39 MFI n = 6) (c *P = 0.038, **P = 0.002, ^#P = 0.04, ^##P = 0.006 for CD39 mRNA and *P = 0.04 for CD39MFI; d *P = 0.049, **P = 0.0028, ^#P = 0.012, ^¶P = 0.01 for CD39 mRNA and *P = 0.049, **P = 0.009, ^#P = 0.018, ^¶P = 0.011 for CD39 MFI using one-way ANOVA followed by Tukey’s multiple comparisons test). Silencing of NCL, alone or in combination with HNRNPA1, resulted in increased CD39 mRNA levels and MFI in Treg and Th17 cells obtained from Crohn’s patients. e Mean ± SEM percentage inhibition of CD4⁺CD25⁻ cell proliferation in the presence of untreated and siRNA (or scramble) treated Treg in HS (n = 7) and Crohn’s disease patients (n = 7) (*P = 0.029, ^#P = 0.032, ^¶P = 0.014, and ^ƒP = 0.037 using one-way ANOVA followed by Tukey’s multiple comparisons test). Silencing of NCL, HNRNPA1, or both enhanced the suppressive function of Treg obtained from patients with Crohn’s disease.