Fig. 6

Leukemic differentiation stage does not affect miR-182 promoter methylation status. A 94 AML samples were divided in CD11b⁺ or CD11b⁻ cells. The frequencies of AML cells with miR-182 promoter hypermethylation and AML cells with hypomethylation were analyzed in CD11b⁺ or CD11b⁻ AML cells. B miR-182 promoter methylation frequency was analyzed in CD11b⁺ or CD11b⁻ AML cells. C CD11b and CD14 staining were performed by flow cytometer in U937 cells treated with 1 μM ATRA, 0.1 μM PMA, or DMSO (1:1000) as control (Ctrl) for 72 h. The representative plots (left) and statistical analysis of CD11b⁺ or CD14⁺ cells were shown (right). D Wright‒Giemsa staining was performed in U937 cells treated with 1 μM ATRA, 0.1 μM PMA, or Ctrl for 72 h. E Bisulfite-genomic sequencing was used to assess the methylation frequency of miR-182 promoter in U937 cells treated with 1 μM ATRA, 0.1 μM PMA, 5 μM DAC, 5 μM AZA, or Ctrl for 72 h. Each row of the circle represents an individual clone. Empty and black circles represent unmethylated and methylated CpG dinucleotides, respectively. The statistical analysis of methylation frequency is shown. ***P < 0.001; ****P < 0.0001. ns: Not significant