Fig. 3

Effects of forced expression of miR-9 on leukemogenic potential of EVI1^high cell lines in vitro and in vivo in EVI1^high leukemia xenograft mice. a Cell cycle status of EVI1^high and EVI1^low leukemia cells with ectopic expression of miR-9 was determined by flow cytometry. Histograms depict the frequency of cells in G0/G1, S and G2 phases of cell cycle. b Representative flow cytometric apoptosis of EVI1^high and EVI1^low with ectopic expression of miR-9. c Histograms depict the frequency of apoptosis in EVI1^high and EVI1^low with ectopic expression of miR-9. d Colony-forming assay. AML cell lines with ectopic expression of miR-9 were cultured in methylcellulose medium and colonies and counted at 7 days of culture. e Frequency of apoptosis in EVI1^high (AML Sample H6 and H7) and EVI1^low (AML sample H12 and H13) cells with ectopic expression of miR-9. f Histograms showing the colony forming ability of EVI1^high, EVI1^low, primary AML cells with ectopic expression of miR-9 or vector. g Kaplan–Meier analysis of xenograft mice (n = 7) injected with AML1 cells with miR-9-Lego or Lego empty control vector. h Representative flow cytometric analysis of engrafted AML-1 cells (hCD45+) and mouse cells in NSG mice (n = 4 to 5). i Histogram depicts the percent AML-1 cells in bone marrow, spleen and peripheral blood after engraftment. All data are representative of two-to-three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001