Figure 2.

Ex vivo effects of everolimus on leukemic spleen cells in combination with imatinib (IM). (a) Leukemic spleen cells were co-cultured with S-17 stromal cells for up to 35 days. Cells were counted with Trypan blue, and viable cells were maintained. (b) Cells were treated with or without everolimus (Eve, 100 n) and imatinib (1 μ) alone and in combination for 5 days on S-17 cells. DNA contents were assessed (upper panels) and Hoechst/PyroninY cell cycle analysis (lower panels) was performed. (c) Percentages of G₀ population in total acquired cells were compared with dimethylsulfoxide (DMSO) control after 5-day treatment with imatinib (1 μ), everolimus (100 n) or in combination. Graph shows the means±s.d. values of three independent experiments (^*P<0.05 by one-way analysis of variance followed by Bonferroni). (d) In the stromal-culturing system, leukemic spleen cells were treated with DMSO (upper column) or in combination of imatinib and everolimus (lower column) for 5 days. Cells were stained with CD34, CD38, human CD45, propidium iodide, and Annexin-V. Cells were gated for human CD45⁺ and CD34/CD38, and cell viabilities in CD34⁺38⁻ population are shown. Panels show a representative analysis.