... control. For trans-fection, cells were plated in 10 cm dishes at 50 % conflu-ency, and siRNA duplexes (50 nm) were introduced intothe cells using Lipofectamine 2000 (Invitrogen) accordingto the ... followed by afinal resuspension in lysis buffer ( 150 mm NaCl, 50 mmTris ⁄ Cl, pH 7.4, 1% NP-40, 0. 25% sodium deoxycholate,1mm EGTA) containing protease inhibitors for westernblot analysis.Flow ... phosphory-lation of Bcl-2 following a variety of stimuli. Theseinclude paclitaxel-activated Raf-1 [ 15] , paclitexel- orvincristine-induced protein kinase A [16], bryostatin-1-induced mitochondrial localized...