Induction of cell death and reduction of clonogenic survival by SGI-1776 in AML cell lines. (A) Flow cytometry analysis of annexin-FITC/propidium iodide staining of MV-4-11 cells that were either untreated, treated with 0.1% DMSO vehicle alone, 0.1, 0.3, 1, 3, or 10μM SGI-1776 for 24 hours. (B) Graphical representation of annexin/PI positivity in MV-4-11 (squares), MOLM-13 (triangles), and OCI-AML-3 (circles) cells treated with 0.1% DMSO vehicle alone, 0.1, 0.3, 1, 3, or 10μM SGI-1776 for 24 hours. The results represent an average of triplicate experiments ± SEM (C-E) Suppression of clonogenic survival of MV-4-11 cells by SGI-1776. Clonogenic assay were performed as described in “Clonogenic assay.” (C) Representative wells of MV-4-11 colonies cultured in drug-free methycellulose after SGI-1776 washout. (D) Images of formed colonies at 10× magnification. Note that the colonies are more densely populated in vehicle DMSO alone than with SGI-1776 treatment. (E) The average percentage of colonies formed relative to DMSO of 3 independent experiments ± SEM.