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. 2021 Nov 25;40:373. doi: 10.1186/s13046-021-02186-0

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© The Author(s) 2021

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — DNMT3A mediates hypermethylation of ELP5. a RT-qPCR analysis of ELP5 transcription levels upon DNMT1, DNMT3a, and DNMT3B stably knockdown in NOZ and GBC-SD cells. b MS-qPCR analysis of ELP5 promoter methylation levels upon DNMT1, DNMT3A, and DNMT3B stably knockdown in NOZ and GBC-SD cells. c ChIP-qPCR analysis of 5mC contents in ELP5 promoter with or without DNMT3A stably silenced in NOZ cells. d Luciferase assay to screen the ELP5 promoter region with the highest pro-transcription activity by transfected with a group of promoter constructs with different 5′ deletions of ELP5 promoter in HEK293T cells. RLU, relative light units. e Luciferase assay of ELP5 promoter (− 450 bp to + 107 bp) activities by ELP5 promoter construct co-transfected with empty vector (EV), wild type (WT), or loss-of-function mutation (R882H) of DNMT3A constructs in HEK293T cells. f-h ChIP-qPCR analysis of 5mC contents in ELP5 promoter (f), RT-qPCR analysis of ELP5 transcription levels (g), and western blot analysis for ELP5 protein levels (h) in both NOZ and GBC-SD cells transfected with EV, WT, or R882H of DNMT3A constructs. Student’s t test for statistical analysis, *P < 0.05, **P < 0.01, ***P < 0.001