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. 2022 May 4;13(5):433. doi: 10.1038/s41419-022-04805-w

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© The Author(s) 2022

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. 3 — A Determination of miR-421 expression in the CRC and adjacent normal tissues (n = 20) by RT-qPCR. B Determination of miR-421 expression in HCT116 and SW480 cells transfected with si-TUG1-1 by RT-qPCR. C The binding of TUG1 to miR-421 in HCT116 cells verified by dual-luciferase reporter assay. D The binding of TUG1 to miR-421 in HCT116 cells tested by RIP assay in combination with qPCR. E The binding of TUG1 to miR-421 in HCT116 cells tested using RNA pull-down. F Determination of KDM2A expression in the CRC and adjacent normal tissues (n = 20) by RT-qPCR. G The binding between miR-421 and KDM2A in HCT116 cells identified using dual-luciferase reporter assay. H The binding of miR-421 to KDM2A in HCT116 cells tested by RIP assay in combination with qPCR. I The binding of miR-421 to KDM2A in HCT116 cells tested using RNA pull-down. J Determination of TUG1, miR-421 and KDM2A expression in HCT116 cells transfected with miR-421 mimic by RT-qPCR. K Measurement of KDM2A protein level in HCT116 cells transfected with miR-421 mimic by western blot assay. L Determination of TUG1, miR-421 and KDM2A expression in HCT116 cells transfected with si-TUG1-1 or combined with miR-421 inhibitor by RT-qPCR. M Measurement of KDM2A protein level in HCT116 cells transfected with si-TUG1-1 or combined with miR-421 inhibitor by western blot assay. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001 and ^****p < 0.0001, compared with adjacent normal tissues, HCT116 cells transfected with si-NC, mimic NC or si-NC + inhibitor NC, or HCT116 cells incubated with IgG or Bio-NC. ^#p < 0.05, ^##p < 0.01, and ^####p < 0.0001, compared with HCT116 cells transfected with si-TUG1-1 + inhibitor NC. The measurement data are expressed as mean ± standard deviation. The experiments were repeated three times independently. Comparison between two groups was conducted by unpaired t-test. Multi-group comparison was conducted by one-way ANOVA with Tukey’s post hoc test.