Am16708 289481

As it was previously described [77 (link)], siRNAs directed against ERRα (Invitrogen, AM16708/289481) with the sense sequence 5′-CCGCUUUUGGUUUUAACC-3′ and antisense sequence 5′-GGUUUAAAACCAAAAGCGG-3′ or control scrambled siRNAs (Invitrogen, AM4611, negative control) were transfected into MCF-7 and MDA-MB-231 cells using Lipofectamine RNAiMAX Transfection Reagent (Invitrogen, Burlington, ON, Canada) following the manufacturer’s instructions. After 48 h post-transfection, fresh phenol red-free medium containing 2% lipoprotein-depleted and charcoal-stripped serum was added, and cells were treated with cholesterol (5 µM for MDA-MB-231, and 10 µM for MCF-7 cells). The knocked-down ERRα breast cancer cells were used for metabolic assays.

siRNAs directed against ERRα (AM16708/289481, Invitrogen, Carlsbad, CA, USA) with the sense sequence 5′-CCGCUUUUGGUUUUAACC-3′ and antisense sequence 5′-GGUUUAAAACCAAAAGCGG-3′ or control scrambled siRNAs (AM4611, negative control, Invitrogen) were transfected into MCF-7 and MDA-MB-231 cells using Lipofectamine RNAiMAX Transfection Reagent (Invitrogen) according to manufacturer’s instructions. At 48 h post-transfection, fresh phenol red-free medium containing 2% lipoprotein-depleted serum was added and cells were treated with cholesterol and/or lovastatin (5 μM for MDA-MB-231, 10 μM for MCF-7 cells). The knock-down breast cancer cells were used for cell growth assays, immunoblotting and qPCR analyses.