Cell lines, cell culture, and transfections

HeLa cells were obtained from the American Type Culture Collection (Manassas, USA) and were cultured in DMEM media (Invitrogen) supplemented with 10% heat inactivated fetal bovine serum (Gibco) and 1% penicillin streptomycin (Corning, Manassas, USA). Cells were cultured in a 5% CO₂ incubator at 37°C.

DsiRNA and antisense oligonucleotide (ASO) transfections were performed using Lipofectamine RNAiMax (Invitrogen) according to the manufacturer's suggested protocol with 48 h incubation. The following dsiRNAs were obtained from Integrated DNA Technologies (Coralville, USA): scaRNA2 sense (5′-rUrGrArUrUrArUrCrGrArGrGrCrGrArUrUrCrUrGrArUrCTG-3′), anti-sense (5′-rCrArGrArUrCrArGrArArUrCrGrCrCrUrCrGrArUrArArUrCrArGrU-3′), scaRNA17 sense (5′-rCrCrGrCrArGrUrArUrUrUrUrCrCrUrUrArUrArUrGrArUCA-3′), anti-sense (5′-rUrGrArUrCrArUrArUrArArGrGrArArArArUrArCrUrGrCrGrGrGrC-3′). ASOs were also obtained from Integrated DNA Technologies, and are as follows: snord16 sense control (5′-mU*mC*mA*mG*mC*G*A*C*A*G*T*T*G*C*C*mU*mG*mC*mU*mG-3′), snord16 antisense (5′- mC*mA*mG*mC*mA*G*G*C*A*A*C*T*G*T*C*mG*mC*mU*mG*mA -3′), scaRNA17 (5′- mA*mU*mA*mU*mA*A*G*G*A*A*A*A*T*A*C*mU*mG*mC*mG*mG -3′), snord68 antisense (5′-mA*mU*mG*mU*mG*C*T*T*T*C*A*T*C*A*A*mG*mG*mC*mC*mG-3′), snord111B antisense (5′-mG*mA*mA*mG*mA*G*A*C*A*T*G*A*A*T*C*mU*mG*mA*mA*mU-3′). Note, the m denotes 2′ O-methylated RNA bases and the * denotes phosphorothioate bonds.