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. 2021 May 24;12(6):532. doi: 10.1038/s41419-021-03814-5

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© The Author(s) 2021

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. 3 — A PEPT1 promoter region (±2.5 kb) around 5’ regions adjacent to the TSS. CGI, CpG islands. BSP, bisulfite-sequencing PCR. B DNA methylation data of PEPT1 promoter in cancerous and adjacent non-tumorous tissues from the TCGA database (***P < 0.001). C BSP analysis of PEPT1 CGI in cancerous and adjacent non-tumorous tissues (n = 11). Methylation percentages of the 35 CpG loci in the sequenced region were calculated. 11 pairs of CRC tissues belong to tissues in Fig. 1B (CRC17, 19, 22, 26, 41, 47, 49, 50, 51, 52, 54). The y axis indicates the average methylation percentage of each CpG site calculated from patient tissues with PEPT1 repression. D–E Calculation of methylation percentages of PEPT1 promoter after DAC and siDNMT1 treatment in CRC cells. F Luciferase assay in DNMT1 knockdown CRC cells. Mock, cells transfected with negative control siRNA and PEPT1 promoter constructs. Si-DNMT1, cells transfected with siDNMT1#2 and PEPT1 promoter constructs. Data are shown as means ± SD, two-tailed unpaired t-test, *P < 0.05, **P < 0.01.