Ki11502 induces G0/G1 cell-cycle arrest and apoptosis of leukemia cells. (A,C,E) Cell-cycle analysis. EOL-1 (A), MOLM13 (C), or MV4–11 (E) cells (5 × 105/mL) were plated in 6-well plates and cultured with Ki11502. After 24 or 48 hours, cells were harvested and cell-cycle distribution was analyzed by FACScan. The numerical results represent the mean of triplicate plates, and a representative experiment is shown. (B,D,F) Annexin V binding. EOL-1 (A), MOLM13 (C), or MV4–11 (E) cells (5 × 105/mL) were plated in 24-well plates and cultured with various concentrations of Ki11502. After 48 hours, cells were harvested and stained for annexin V and propidium iodide (PI), and cells were analyzed by FACScan. Lower left quadrants: viable cells. Lower right quadrants: early apoptotic cells (annexin V+, PI−). Upper right quadrants: nonviable, late apoptotic/necrotic cells (annexin V and PI+). The numerical results represent the mean of triplicate plates, and a representative experiment is shown. (G) Western blot analysis. EOL-1, MV4-11, and MOLM13 cells were cultured with various concentrations of Ki11502 (0.1-1 nM for EOL-1 and 0.1-1 μM for MV4-11 as well as MOLM13). After 48 hours, cells were harvested, and proteins were extracted and subjected to Western blot analysis. The membranes were sequentially probed with anti–Bcl-2, Bcl-xL, Mcl-1, and anti–β-actin antibodies. Results represent one of the 3 experiments performed independently, each giving similar results.
