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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. 2 — AP4 directly represses MIR22HG. A qPCR analysis of MIR22HG and p21 in the indicated clones of DLD-1 cells. B qPCR analysis of miR-22-3p expression (left panel) and nascent MIR22HG mRNA (right panel). C qPCR analysis of MIR22HG expression after AP4 induction by DOX for the indicated duration (left panel) and nascent MIR22HG 48 h after AP4 induction by DOX (right panel). D qPCR analysis of MIR22HG (left panel) and miR-22-3p (right panel) after c-MYC induction by DOX for indicated durations. E qPCR analysis of nascent MIR22HG 48 h after c-MYC induction by DOX. F ChIP-Seq enrichment profiles for AP4-VSV associated chromatin were generated with the UCSC genome browser. The gene structure ideogram is shown below the ChIP-seq tracks. G qChIP analysis of AP4 occupancy. 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. H qPCR analysis of MIR22HG in the indicated cells 48 h after transfections. I miR-22-3p anti-sense reporter dual-luciferase analysis 48 h after transfection. J β-gal detection at pH 6 48 h after transfection. Quantification of 3 fields with 120 cells in total. Scale bars: 50 μm. K Detection of γH2AX foci 48 h after transfection. Quantification of 3 fields with 120 cells total. Scale bars: 20 μm. In panel A-E and G-K the mean + SD (n = 3) is provided with *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001