Co-treatment of leukemic cell lines with embelin and LY294002 potentiated apoptosis.

<p>K562 <b>(A)</b> and U937 <b>(B)</b> cells were treated either with 5 μM embelin or 5 μM LY294002 alone or in combination for 24 h and cell proliferation assays were performed using MTT as described in Materials and Methods. The graph displays the mean +/- SD of three independent experiments for all the doses. <b>*</b><i>P</i> < 0.001. Subtoxic doses of Embelin and LY294002 potentiated apoptosis in leukemic cells. K562 <b>(C)</b> and U937 <b>(D)</b> cells were treated either 5 μM embelin or 5 μM LY294002 alone or in combination for 24 h, cells were subsequently stained with fluorescein-conjugated annexin-V antibody and propidium iodide (PI) and apoptosis was analyzed by flow cytometry. Percentage of apoptotic cells (early + late apoptosis) were calculated. The graph displays the mean +/- SD (standard deviation) of three independent experiments. Subtoxic doses of embelin and LY294002 potentiated the activation caspases in leukemic cells. K562 <b>(E)</b> and U937 <b>(F)</b> cells were treated either with 5 μM embelin or 5 μM LY294002 alone or in combination for 24 h. Cells were lysed and equal amount of proteins were immunoblotted with antibodies against XIAP, p-Akt, Akt, cleaved caspase-3 and PARP. GAPDH was used to measure equal loading.