Non targeting pool

Experiments were carried out 48 h after transient transfection (Amaxa Nucleofector, Lonza, Basel, Switzerland) with siRNA oligonucleotides (ON-TARGETplus Human RIP1 siRNA SMART pool and NON-TARGETING pool; Dharmacon, Lafayette, USA). REH cells with stable (shRNA) RIP1 knockdown were kindly provided by S Löder and Dr. Fulda, Ulm, Germany.^{29 (link)}

Glioma cells were transiently transfected with 100 μM of NFкB (RelA)-specific siRNA (ON-TARGETplus Human RelA siRNA-SMARTpool, Dharmacon) and 100 μM control-siRNA (ON-TARGETplus Non-targeting Pool) for 48 h followed by mRNA expression and promoter-activity analysis in pGL3F1 co-transfected cells. Cell migration/invasion assay and anchorage-independent growth assays were performed after knockdown of NFкB (RelA) following the methods previously published [19 (link)].

Non-targeting siRNA-pool (ON-TARGETplus Non-targeting Pool, # D-001810-10-05) and Mcl-1 (SMARTpool: ON-TARGETplus Mcl-1 siRNA, L-004501-00-0005) were purchased from Dharmacon and transfected as previously described^{21 (link), 35 (link)}. For single siRNA experiments involving Mcl-1, the following sequences were employed: siRNA-1: GGU UUG GCA UAU CUA AUA A; siRNA-2: GAA GGU GGC AUC AGG AAU G, siRNA-3: GAU UAU CUC UCU CGG UAC CUU, siRNA-4: CGA AGG AAG UAU CGA AUU U (Dharmacon). For siRNA experiments, involving Bcl-xL, siRNAs were either used from Ambion (Silencer^® Select s1920; AUA CUU UUG UGG AAC UCU ATT) or Cell Signaling Technology^® (Signal silence^® Bcl-xL siRNA I; #6362). Silencing of PMAIP1 was performed using Silencer^® Select siRNA-1 (s10708; AGA UAU GAA UGU UUC UAA ATT) and siRNA-2 (s10709; AGU CGA GUG UGC UAC UCA ATT) from Ambion. BAK1 knock-down was performed using Silencer^® Select s1880 from Ambion. Briefly, cells were incubated for 6 h with the formed complexes of Oligofectamine^® 2000 (Invitrogen, Carlsbad, CA) and the respective siRNA (12-well condition) in DMEM without FBS and antibiotics. After 6 h, FBS was added to a total concentration of 1.5%.