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. 2022 May 27;21:120. doi: 10.1186/s12943-022-01581-1

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© The Author(s) 2022

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. 4 — MDC1 is directly regulated by AP4. A qPCR analysis of nascent MDC1 mRNA 48 h after AP4 induction by DOX (left panel). qPCR analysis of nascent MDC1 mRNA in untreated cells (right panel). B ChIP-Seq analysis of AP4 occupancy using the UCSC genome browser. C Validation of MDC1 as AP4 direct target via qChIP assay. Cells were treated with DOX for 48 h. Chromatin was enriched by anti-AP4 or anti-rabbit-IgG antibodies. SNAI1 and 16q22 served as positive and negative controls, respectively. D-E qPCR and Western blot analysis of MDC1 in the indicated cell lines 48 h after transfection. F Western blot analysis of MDC1 after AP4 induction by DOX for the indicated durations. G qPCR analysis of nascent MDC1 mRNA after c-MYC induction by DOX for 48 h. H Western blot analysis after c-MYC induction by DOX for the indicated periods. I Model of the AP4/miR-22-3p/MDC1 coherent feed-forward loop. In panels A, C, D and G, the mean + SD (n = 3) is provided with *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001