MB1-47 antileukemic effects in T-ALL cell lines in vitro. (A) Relative cell survival of 6 independent human T-ALL cell lines (including PTEN⁺ and PTEN⁻ cells) in the presence of MB1-47 at the indicated concentrations for 72 hours (mean ± SD; n = 3). (B) Representative flow cytometry histograms showing cell-size changes in G₁-gated DND41 and JURKAT cells treated with DMSO (control) or MB1-47 (4 or 2 µM, respectively) for 72 hours. (C) Representative flow cytometry plots of annexin V (apoptotic cells) and 7-aminoactinomycin D (7-AAD; dead cells) staining. Numbers in quadrants indicate percentage of cells. (D) Quantification of apoptosis from DND41 triplicates treated for 72 hours with DMSO (control) or 4 μM MB1-47 (mean ± SD). Statistical significance (P) was determined by using the 2-tailed Student t test; ***P < .005. (E-F) Flow cytometry representation (E) and quantification (F) of cell-cycle analysis of DND41 cells treated with DMSO (control) or MB1-47 (4 μM) for 72 hours (n = 3; mean ± SD). P values in panel F were calculated using 2-way analysis of variance (ANOVA) for multiple comparisons. ***P < .005. FSC-H, forward scatter height; PI, propidium iodide.