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. 2019 Nov 25;10:5334. doi: 10.1038/s41467-019-13313-z

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© The Author(s) 2019

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. 2 — REG1CP promotes transcription of REG3A. a–c Silencing of REG1CP (a) decreased REG3A mRNA (b) and protein levels (c). d Co-introduction of a shRNA-resistant REG1CP mutant (REG1CP-R) diminished the inhibitory effect of REG1CP silencing on REG3A mRNA expression. e, f Overexpression of REG1CP increased REG3A mRNA (e) and protein (f) levels. g, h Silencing of REG1CP decreased (g) whereas overexpression of REG1CP (h) increased enrichment of H3K4me3 to REG3A promoter, but not to REG1A or REG1B promoter (g). Relative enrichment of H3K4me3 to the -7/-109 fragment of REG3A promoter was measured using ChIP-qPCR assays in LIM1215 and HT-29 cells after silencing (g) or in SW480 and COLO205 cells after overexpression (h) of REG1CP. i, j Silencing of REG1CP increased (i) whereas overexpression of REG1CP decreased (j) enrichment of H3K27me3 to REG3A proximal promoter. Relative enrichment of H3K27me3 at the -7/-109 fragment of REG3A promoter was measured using ChIP-qPCR assays in LIM1215 and HT-29 cells after silencing (i) or in SW480 and COLO205 cells after overexpression (j) of REG1CP. k, l Regression analysis of the relationship between REG3A mRNA and REG1CP expression in two cohorts of colon cancer samples determined using qPCR (k) and in published datasets acquired from the R2 Genomics Analysis and Visualization Platform (l). n = 3 independent experiments unless specified differently. Data are presented as Mean ± SEM (a, b, d, e, g–j) or representatives (c, f). Statistical significance was calculated using a two-tailed t-test unless specified.