Fig. 4

Combination treatment with sub-optimal doses of embelin and LY294002 synergistically induces apoptosis in BC cells. (a) AKT siRNA down-regulates expression of XIAP in BC cells. MDA-MB-231 cells were transfected with siRNA targeted against AKT and proteins were isolated and probed with antibodies against p-AKT, AKT, XIAP and GAPDH. (b and c) Combination of embelin and LY294002 at sub-optimal doses synergistically inhibits cell viability and induces apoptosis in BC cells. EVSAT and MDA-MB-231 cells were treated with either alone or in combination of embelin (10 μM) and LY294002 (10 μM) for 24 h. Following 24 h treatment, cells were analysed for cell viability by MTT assays (b) and apoptosis after staining the cells with annexin V/PI by flow cytometry (c). (d) Combination treatment causes inactivation of AKT, down-regulation of XIAP and caspase-dependent apoptosis in BC cells. EVSAT and MDA-MB-231 cells were treated with combination of sub-toxic doses of embelin and LY294002 for 24 h. Following incubation, proteins were isolated and probed with antibodies against XIAP, p-AKT, AKT, Bcl-Xl, caspase-9, caspase-3, PARP and GAPDH