Fig. 3: Mir139 is epigenetically silenced by PRC2 in MLL-AF9 cells.

A ChIP-seq reads of the Pde2a locus (chr7:108,596,060–108,645,660) are depicted. The top two tracks show H3K4me3 (red) and H3K27Ac (orange) enrichment in common myeloid progenitors (CMPs). The next track indicates RNA Polymerase II (POL-II) binding (purple) of MLL-AF9 cells. The next three tracks depict H3K27me3 (blue), H3K27Ac and SUZ12 (green) enrichment in MLL-AF9 cells treated with UNC2400 (inactive PRC2 inhibitor). The bottom three tracks show the same data of MLL-AF9 cells treated with UNC1999 (active PRC2 inhibitor). Scale bar indicates 5 kilobases (kb). Black bars indicate exons. B The methylation status of CpGs in the PDE2A promoter region, enhancer region 1 (E1) and E2 of THP-1 and Molm-13 cells is depicted. Closed blue circles indicate methylated CpG, open blue circles indicate unmethylated CpG. C Schematic overview of the Pde2a locus (chr7:108,596,060–108,645,660). Fold enrichment of H3K27me3 (orange) and control IgG (white) antibody binding to the indicated regions in MLL-AF9 cells treated with depicted concentrations of UNC1999 as determined by ChIP-qPCR is shown. Arrows indicate the primer sets. The scale bar indicates 2.5 kb. D Viability of WT or Mir139KO MLL-AF9 cells treated with UNC1999 relative to mock-treated WT or Mir139KO MLL-AF9 cells is shown. Data are representative of three experiments. E Expression levels of miR-139-3p and miR-139-5p in MLL-AF9 cells treated with UNC1999 relative to snRNA U6 and untreated MLL-AF9 cells are shown. Data are representative of three experiments. F Viability of Molm-13 cells treated with UNC1999 relative to mock-treated Molm-13 cells, is shown. Depicted data are representative of three experiments. G Expression levels of miR-139-3p and miR-139-5p in Molm-13 cells treated with UNC1999 relative to snRNA U6 and untreated Molm-13 cells are shown. Depicted data are representative of three experiments. H Viability of UNC1999-treated primary MLL-AF9 patient samples (n = 3) relative to untreated cells is shown. I Expression levels of miR-139-5p and miR-139-3p in primary MLL-AF9 patient samples (n = 3) treated with UNC1999 relative to snRNA U6 and untreated cells are shown. All graphs show mean ± SEM. The two-tailed unpaired student’s t test was used in C–E and G. The two-tailed paired student’s t test was used in F, H and I.