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. 2021 Jun 13;22:180. doi: 10.1186/s13059-021-02402-2

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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. 5 — Experimental validation of m⁶A on processed pseudogene DSTNP2 affects the expression of DSTN in GM12878 cell line. a Relative gene expression of DSTNP2 and DSTN after DSTNP2 knockdown using siRNA. * P< 0.05; *** P< 0.001 (two-tailed t test). b Western blot comparing the protein levels of DSTN in control and DSTNP2 knockdown cells. c The relative expression of DSTN upon overexpression empty vector and wild type DSTNP2 respectively. Error bars represent standard errors. *** P< 0.001 (two-tailed t test). d The relative expression of DSTN at different time points after transcription inhibition in control and DSTNP2 knockdown cells respectively. Error bars represent standard errors. * P< 0.05; ** P< 0.01; *** P< 0.001 (two-tailed t test). e The relative expression of DSTN and DSTNP2 after adding control inhibitor versus miR362-5p inhibitor. f Bar plot comparing the relative expression of total, cytoplasmic and nuclear DSTN respectively upon overexpression of wild type versus m⁶A sites-mutated DSTNP2. g Western blot comparing the protein levels of DSTN upon overexpression of wild type DSTNP2 versus m⁶A site-mutated DSTNP2. h, i The relative expression of DSTNP2 (h) and DSTN (i) upon overexpression of wild type DSTNP2 versus m⁶A site-mutated DSTNP2 at different time points after transcription inhibition. Error bars represent standard errors. ** P < 0.01; *** P < 0.001 (two-tailed t test). j The relative expression of DSTN upon overexpression of wild-type DSTNP2 versus m⁶A site-mutated DSTNP2 after adding control inhibitor and miR362-5p inhibitor respectively. Error bars represent standard errors. * P < 0.05 (two-tailed t test). k The relative expression of DSTN at different time points after transcription inhibition upon overexpression of wild-type DSTNP2 versus m⁶A site-mutated DSTNP2 after adding control inhibitor and miR362-5p inhibitor respectively. Error bars represent standard errors. ** P< 0.01; *** P< 0.001. l Western blot showing the protein level of DSTN upon overexpression of wild type DSTNP2 versus m⁶A site-mutated DSTNP2 after adding control inhibitor and miR362-5p inhibitor, respectively