Fig. 5

Transcriptome-wide identification of FTO targets in t(8;21) AML. A Western blot analysis of the expression of FTO in Kasumi-1 cells overexpressing or not wild-type FTO. B The predominant consensus motif RRACH (R[G/A]; H[U/A/C]) analyzed by HOMER in m⁶A-seq (p = 1e−10823). C Density distribution of m⁶A peaks across the length of mRNA transcripts. Each region of 5′-UTR, coding region (CDS), and 3′-UTR was split into 100 segments, and the percentage of m⁶A peaks that fall within each segment was determined. D Distribution of peaks with a significant change in both the m⁶A and RNA levels in FTO-overexpressing Kasumi-1 cells compared with control Kasumi-1 cells. E Venn diagram for the overlapping genes enriched in m⁶A hypomethylated genes in m⁶A-seq and FTO-expression-correlated genes in TCGA-AML database. F Confirmation of changes in the levels of m⁶A of 7 representative m⁶A-hypo genes and 2 non-m⁶A-hypo genes in Kasumi-1 cells by gene-specific m⁶A qPCR. G Kaplan–Meier analysis of event-free survival (left) and overall survival (right) of patients with de novo t(8;21) AML (n = 26) based on the expression of IGFBP2. H Comparison of IGFBP2 mRNA expression in BM samples from AML patient with high versus poor response to Ara-C (GSE97393). I Pearson correlation of the expression of IGFBP2 and FTO in BM cells from 26 patients with de novo t(8;21) AML. J The m⁶A level in IGFBP2 mRNA transcripts in FTO-overexpressing and control Kasumi-1 cells as detected by m⁶A-seq. The m⁶A peaks shown in red rectangles reveal a significant decrease in abundance in FTO-overexpressing compared with mock cells (fold change = 0.72, p = 0.001). The abundance of m⁶A peaks after input normalization are shown. K The expression level of IGFBP2 mRNA in FTO-overexpressing and control Kasumi-1 cells as detected by RNA-seq (fold change = 2.22, p < 0.001)