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. 2007 Nov 19;179(4):701–715. doi: 10.1083/jcb.200703040

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Copyright © 2007, The Rockefeller University Press

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Figure 7. — siRNA silencing of Mcl-1 sensitizes Bak-proficient DU145 cells for Nbk-induced apoptosis and cytochrome c release. Mcl-1 was down-regulated in DU145 mock and Bak cells by use of siRNA. 24 h later, cells were transduced with Ad-mycNbk-tTA to induce Nbk expression and cultured for additional 24 h under off or on conditions. (A) SiRNA silencing of Mcl-1, overexpression of Nbk, and the Bax expression status were confirmed by Western blot analysis. (B) The percentage of apoptotic cells was determined by flow cytometric measurement of hypodiploid DNA. Data are means ± SD from three independent experiments. (C) Cytosolic extracts were prepared and subjected to immunoblotting to analyze Nbk-induced cytochrome c release after down-regulation of Mcl-1.