Fig. 1

High EVI1 expression in AML cell lines and primary AML BM cells from patients is associated with hypermethylation of the miR-9 promoter. a EVI1 expression determined by western blot in AML cell lines. b DNA methylation profile of CpG dinucleotides in the promoter region of miR-9 in AML cell lines. The cycles represent the average methylation level of a specific CpG dinucleotide. Unmethylated, partially methylated, and fully methylated are indicated by open circles, partially blacked circles and blacked circles c Increased methylation in EVI1^high cell lines depicted using histogram plot. The y axis represents the percent methylation of the miR-9 promoter. d Histogram showing average percent methylation of the CpG islands of the miR-9 promoter in the AML cell lines before and after treatment with 5-AZA. Methylation was assessed using bisulfite conversion and direct sequencing of cell lines treated with 5-AZA for 48 h. e Histogram showing fold increase in miR-9 expression in EVI1^high cell lines AML-1 and Kasumi-3 relative to EVI1^low cell line, U-937 after 5-AZA treatment. f qRT-PCR analysis of miR-9 in pediatric primary AML BM cells. g Left panel: Methylation profile of promoter region of miR-9 in pediatric AML patient BM cells with EVI1^high (AMLH1-H7), and EVI1^low (AML H8-H13) expression. Right panel: Increased percent methylation in EVI1^high patient samples depicted using histogram. The y axis represents the percent methylation of the miR-9 promoter. All data are representative of two-to-three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001