Overexpression of XIAP inhibits TQ and TAM-induced apoptosis.
(A) MCF-7 and MDA-MB-231 cells were transfected with pcDNA3-XIAP-Myc construct. Cell lysates were subjected to western blotting analysis using XIAP, p-Akt, caspase-9, PARP, and β-Actin (Equal loading marker) antibodies. (B) Densitometric analysis of XIAP, p-Akt, caspase-9 and PARP protein levels in transfection blot where each bar represents three independent experiments; P<0.05 (t-test). Light and dark color bars represent pcDNA3.1 and pcDNA3-XIAP-Myc transfected cells, respectively. (C) Early and late apoptosis of pcDNA-XIAP-Myc cotransfected cells were analysed through flow cytometer after staining the cells with Annexin V-FITC and PI. (D) MCF-7 and MDA-MB-231 cells were cotransfected with siRNA-XIAP and pcDNA3-XIAP-Myc construct. Cell lysates were subjected to western blotting analysis using XIAP. (E) Densitometric analysis of XIAP protein levels in cotransfection blot where each bar represents three independent experiments; P<0.05 (t-test). (F) At the end of the treatment period, cells were collected from annexin studies (for apoptotic cell count).