Silencer select sirna 1

HeLa cell line was transfected with siRNA against either non‐targeting (Mission siRNA Universal negative control SIC001 from SIGMA) or SSU72 (Silencer Select siRNA‐1 (ID s26487) and Silencer Select siRNA‐2 ID (s223816) from Ambion) using mission siRNA transfection reagent (Sigma S1452) at 10 nM following manufacturer's instructions. 3 days after transfection, cells were fixed, and IF‐FISH was carried out to quantify telomere dysfunction.

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%.