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Overexpression of XIAP inhibits TQ and TAM-induced apoptosis.

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posted on 2013-04-17, 01:20 authored by Shashi Rajput, B. N. Prashanth Kumar, Siddik Sarkar, Subhasis Das, Belal Azab, Prasanna K. Santhekadur, Swadesh K. Das, Luni Emdad, Devanand Sarkar, Paul B. Fisher, Mahitosh Mandal

(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).

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