Azacitidine induces hypomethylation and changes in CTCF occupancy. (A) Density plots for β-values of all normalized CpGs (n = 723 768) in azacitidine and mock-treated AML samples. (B) MA plot of differentially bound CTCF sites between azacitidine- and mock-treated samples (FDR <0.001). Dark pink dots show peaks with fold change >1.5. (C) PCA analysis of differentially methylated sites (left), differentially bound sites (middle), and differentially expressed genes (right) based on EPIC array data, ChIP-seq for CTCF, and RNA-seq, respectively, in azacitidine- vs mock-treated AML samples. (D) Dot plot of DBCs in relation to changes in gene expression between azacitidine- vs mock-treated samples. The x-axis represents fold change in gene expression and the y-axis fold change in CTCF occupancy (FDR <0.001). Black dots represent genes with statistically significant changes in gene expression (FDR <0.05, fold change >1.5); χ² test was used to calculate statistical significance between up- and downregulated genes (P < .001). (E) Dot plot of DBCs in relation to fold change in DNA methylation by β-values (Δβ-value) for azacitidine- vs mock-treated samples. The x-axis represents Δβ-value (FDR<0.1) and the y-axis fold change in CTCF occupancy (FDR <0.001). Green and red dots represent up- or downregulated genes, respectively (FDR <0.05, fold change >1.5); χ² test was used to calculate statistical significance between Q1 and Q2 (P < .001).