Fig 2: MIR670HG upregulates DNA methylation level at CD24 promoter via m6A modification, FXR1 and TET1. A Schematic of human CD24 and mouse Cd24a genomic loci. Green boxes indicate CpG islands. B Bisulfate DNA sequencing analysis of CpG236 in SW620 cells with MIR670HG overexpression or control. C Bisulfate DNA sequencing analysis of CpG115 in Hepa1-6 cells with Mir670hg knockdown or control. D CUT & RUN assays detecting the binding of TET1 to CD24 promoter in SW620 cells with MIR670HG overexpression or control. E CUT & RUN assays detecting the binding of TET1 to Cd24a promoter in Hepa1-6 cells with Mir670hg knockdown or control. F MeRIP assays detecting m6A modification level of wild-type or 772 site mutated MIR670HG in SW620 cells with wild-type (WT) or 772 site mutated (Mut) MIR670HG overexpression. G MeRIP assays detecting m6A modification level of wild-type or 646 site mutated Mir670hg in MC38 cells with wild-type or 646 site mutated Mir670hg overexpression. H RIP assays detecting the binding of FXR1 and TET1 to wild-type or 772 site mutated MIR670HG. I CUT & RUN assays detecting the binding of TET1 to CD24 promoter in SW620 cells with wild-type or 772 site mutated MIR670HG overexpression. J Bisulfate DNA sequencing analysis of CpG236 in SW620 cells with wild-type or 772 site mutated MIR670HG overexpression. K qPCR analysis of CD24 expression in SW620 cells with wild-type or 772 site mutated MIR670HG overexpression. L Flow cytometry analysis of CD24 protein expression in SW620 cells with wild-type or 772 site mutated MIR670HG overexpression. M qPCR analysis of Cd24a expression in MC38 cells with wild-type or 646 site mutated Mir670hg overexpression. N Flow cytometry analysis of CD24 protein expression in MC38 cells with wild-type or 646 site mutated Mir670hg overexpression. Results are shown as mean ± SD of n = 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns, not significant, by Student’s t-test (B, D, F-H) or one-way ANOVA followed by Dunnett’s multiple comparisons test (C, E, I-N)