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. 2020 Jul 23;11(7):566. doi: 10.1038/s41419-020-02795-1

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© The Author(s) 2020

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. 4 — a A schematic showing potential m6A motifs in regions of PML-RARα mRNA. MeRIP assays were performed in NB4 cells after knockdown of m6A methyltransferases, MALAT1 or a negative control with siRNAs. Then, PCR sequencing for these sites, including potential motifs, were carried out; agarose electrophoresis was shown. b Schematic diagram of METTL14-methylated m6A sites in PML-RARα mRNA exon 4. c PML-RARA mRNA dispersion in nuclei and cytoplasm was detected after reducing the m6A level by knocking down the methyltransferases. d RNA FISH analysis showing the PML-RARα mRNA location in cells by two probes, Cy3-PML (red) and FITC-RARα (green). Scale bar represents 4 μm. e The protein levels of PML-RARα, METTL3, METTL14, and WTAP were monitored after knockdown of m6A methyltransferases by several different siRNAs. Three independent experiments were carried out. f Schematic diagram showing RNA FISH assays for testing PML-RARα mRNA transport by transfecting PML-RARα-EGFP-12×MS2bs and YTHDC1-MS2 constructs with knockdown of m6A in HEK-293T cells. g Representative images showed fusion protein green dots were reduced. Red dots were remained in nucleus when m6A methyltransferases were knocked down in HEK-293T cells. Scale bar represents 4 μm. h Combined with cotransfecting YTHDC1 into HEK-293T cells, representative images of PML-RARα mRNA and PML-RARα-EGFP are shown in HEK-293T cells after reducing m6A levels. Scale bar represents 4 μm. i Combined with cotransfecting YTHDC1-MS2, representative images showed PML-RARα mRNA retention in nuclei was significantly restored after reducing m6A, as compared with the negative controls. Scale bar represents 4 μm. j Schematic diagram showing the hypothesis the potential roles of MALAT1, m6A, and the reader YTHDC1 in the fusion gene export process.