To carry out the siRNA experiment, an adequate (2 × 105 cells/well) number of cells (RAW 264.7 cells) were first cultured in 6-well plates for 24 h and allowed to grow until ~60% confluent. Then cells were transfected with 10–50 nM siRNA using Lipofectamine RNAiMax (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. si-Control RNA and si-Nrf2 RNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Si control rna
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Si-Control RNA is a synthetic RNA molecule used as a control in RNA interference (RNAi) experiments. It serves as a negative control to assess the specificity of gene silencing effects observed with small interfering RNA (siRNA) or short hairpin RNA (shRNA) treatments. Si-Control RNA does not target any known gene and thus provides a baseline for comparison to evaluate the efficacy and off-target effects of experimental RNAi.
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3 protocols using si control rna
1
RAW 264.7 Cell siRNA Transfection
2
Nrf2 Silencing in Cell Line
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Cells (1 × 105 cells/mL) were transfected with 10–50 nM siRNA using Lipofectamine RNAiMax (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. si-Control RNA and si-Nrf2 RNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
3
RNAi-Mediated Knockdown of NRF2
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Lipofectamine® RNAiMax (cat. no. 13778075; Thermo Fisher Scientific, Inc.) was used to transfect 20 nM siRNA against NRF2 (siNRF2 RNA) (cat. no. sc-37030; Santa Cruz Biotechnology, Inc.) or negative control (siControl RNA) (cat. no. sc-37007; Santa Cruz Biotechnology, Inc.) into cells. The siRNA sequences were as follows: Control siRNA sense, 5'-CACAGGGUAAGGAACUCGUCUCUCA-3' and antisense, 5'-UGAGAGACGAGUUCCUUACCCUGUG-3'; and NRF2 siRNA sense, 5'-GCAUGCUACGUGAUGAAGAtt-3' and antisense, 5'-UCUUCAUCACGUAGCAUGCtt-3'. After incubation for 24 h at 37˚C, the transfected cells were processed for ROS detection and β-galactosidase staining assay.
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