Parental cell lines were obtained from ATCC and cultured in DMEM/10% FBS (Hyclone). FR cells were maintained in 1 µM fulv (Tocris Bioscience). MCF-7/FR cells were a gift from M. Ellis (Baylor College of Medicine, Houston, TX). ZR75-1/FR cells were generated by culturing ZR75-1 cells with 1 µM fulv for 4 mo. Cells were transfected with siRNA targeting Bim (Dharmacon, Qiagen), Cyclin D1 (Ambion), or nonsilencing control (Qiagen) using Lipofectamine RNAiMAX (Life Technologies) per the manufacturer’s instructions.
Nonsilencing control
Manufactured by Qiagen
The Nonsilencing control is a laboratory tool used to assess the efficacy of RNA interference (RNAi) experiments. It serves as a control to distinguish between the specific effects of a targeted gene silencing and non-specific effects caused by the RNAi process itself.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using nonsilencing control
1
Fulvestrant-resistant Breast Cancer Cell Lines
2
HuR and miR-21 Regulation in Oxidative Stress
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HeLa cells were maintained in standard conditions in Dulbecco's modified Eagle's medium (DMEM) supplemented with heat-inactivated 10% fetal calf serum, 2 mM L-glutamine, and 1% antibiotics (100 units/ml penicillin-streptomycin). The GST-HuR expression plasmid was described previously [38 (link)]. Transfections of siRNA, or miRVana microRNA mimics and inhibitors were performed using Lipofectamine RNAiMax (Life Technologies). Briefly, 2.5 × 104 HeLa cells were seeded in 12-well plates for 24 hours. Transfections were performed at a final concentration of 20 nM HuR siRNA (AAGUCUGUUCAGCAGCAUUGGUUdTdT, Dharmacon), nonsilencing control (Qiagen, Cat. # 1022076), miR-21 mimic (Ambion, Cat. # 4464066), miR negative Control mimic (Ambion, Cat. # 4464058), anti-miR-21 (Ambion, Cat. # 4464084), anti-miR control (Ambion, Cat. # 4464076). Cells were treated in the presence of 0.5 mM H2O2 for 4 hours. Cells were harvested for analysis after the indicated time points as described below.
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