4.6. Inhibition of PGC-1α Expression by Small Interfering RNA (siRNA)

SNU-C5/5FUR cells were seeded in 60 mm dishes and were transfected with siRNA in serum-free Opti-MEM (Gibco BRL) by using Lipofectamine 2000, following the manufacturer’s instructions (Thermo Fisher Scientific). After transfection with siRNA for 48 h, the cells were treated with 5FU for 24 h. The siRNA used to block PGC-1α (The PGC-1α siRNA no. 1 sequence: 5′-UCACCGAGACCGACGUUAA-3′, no. 2 sequence: 5-GAUCGAGCAUGGUCCUCUU-3′, no. 3 sequence: 5′-AGAUGUGUAUCACCCAGUA-3′, and no. 4 sequence: 5′-GACCGUUACUAUCGUGAAA-3′) and a scrambled sequence (The scrambled siRNA no. 1 sequence: 5′-UGGUUUACAUGUCGACUAA-3′, no. 2 sequence: 5′-UGGUUUACAUGUUGUGUGA-3′, no. 3 sequence: 5′-UGGUUUACAUGUUUUCUGA-3′, and no. 4 sequence: 5′-UGGUUUACAUGUUUUCCUA-3′) were purchased from Dharmacon (Lafayette, CO, USA).