ShRNA transfections were performed using the MegaTran 1.0 Transfection Reagent (OriGene) according to the manufacturer’s instructions. The lentiviruses were used to infect NSCLC cells with Polybrene (8 μg/ml) for 4 h. The original medium was replaced with fresh medium 12 h later. The siRNAs, siNrf2, and siNestin, were purchased from Ribobio, and their encoding vectors were transfected into NSCLC cells using the Lipofectamine RNAiMAX Transfection Reagent (Invitrogen).
Sinrf2
Manufactured by RiboBio
Sourced in China
SiNrf2 is a small interfering RNA (siRNA) targeting the Nrf2 gene. Nrf2 is a transcription factor that regulates the expression of genes involved in the antioxidant response. SiNrf2 is designed to downregulate the expression of Nrf2, thereby modulating the cellular antioxidant defense system.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (6 mentions)
Si nc, by RiboBio (3 mentions)
Megatran 1.0 transfection reagent, by OriGene (1 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (1 mentions)
Sinestin, by RiboBio (1 mentions)
Antagomir 200a, by RiboBio (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Pcdna3.0 plasmid, by Thermo Fisher Scientific (1 mentions)
Sipink1, by RiboBio (1 mentions)
Siparkin, by RiboBio (1 mentions)
Si control, by RiboBio (1 mentions)
Penter vector, by Charles River Laboratories (1 mentions)
Lipofectamine 3000 transfection reagent, by RiboBio (1 mentions)
9 protocols using sinrf2
1
Transfection of NSCLC cells with siRNAs
2
Nrf2 Silencing in Chondrocytes
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First, siRNA negative control (si-Crtl) and its target siRNA Nrf2 (si-Nrf2) by Ribobio (Guangzhou, China) were constructed and synthesized. Then, chondrocytes were seeded in a 6-well plate for 24 h. Transfection with si-Nrf2 or si-NC were carried out using Lipofectamine 2000 reagent according to the protocol.
3
Adenoviral Knockdown of CTRP3 in Adipocytes
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Adenoviral vectors carrying CTRP3 small hairpin RNAs (shRNAs) designed to knock down CTRP3 in adipocytes were generated by Hanbio (Shanghai, China). Scrambled shRNA was used as a control. Briefly, adipocytes were incubated in DMEM/F12 and were infected with adenoviral vectors at a multiplicity of infection (MOI) of 100 for 4 h. To knock down Nrf2 in H9c2 cells, we purchased siNrf2 and negative-control siRNA from Guangzhou RiboBio Co., Ltd. H9c2 cells were transfected with siNrf2 using Lipofectamine RNAiMAX (Invitrogen) for 24 h. Transfection efficiency was confirmed by western blotting. AntagomiR-200a and a miRNA inhibitor control, which were obtained from Guangzhou RiboBio Co., Ltd, were used to silence miR-200a in H9c2 cells.
4
Constructing ALK7-overexpressing Plasmid
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The full-length ALK7 cDNA was cloned into the pcDNA3.0 plasmid (Invitrogen Corporation, San Diego, CA, USA) to construct the ALK7-overexpressing plasmid, pcDNA3.0-ALK7. The mammalian expression vector pcDNA3.0 was used as the transfection control. si-ALK7 and si-Nrf2 were obtained from RiboBio Co. Ltd.. HBZY-1 cells were transfected using lipofectamine 2000 (Invitrogen).
5
Nrf2 Knockdown in Microglia Transfection
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For transfection, microglia were inoculated at a density of 40–60% and infected with lentivirus at a multiplicity of infection (MOI) of 20. After 12 h of transfection, the medium was changed. Following cell confluence, the transfected microglia were passaged for further experiments. To suppress the expression of Nrf2, microglia (50–60% confluence) were transfected with Nrf2 small interfering RNAs (Si-Nrf2; RiboBio, China, Supplementary Table 3 ) according to the manufacturer’s protocol. The transfection efficiency was evaluated by western blot analysis.
6
Overexpression and Silencing of CTRP3 in HK-2 Cells
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Expression plasmids containing CTRP3 cDNA (pcDNA-CTRP3) and negative controls (vector) were constructed by Jennio Biotech (Guangzhou, China). CTRP3 siRNA (siCTRP3) and negative control siRNA (siNC) were designed and synthesized by RiboBio (Guangzhou, China) and transfected to HK-2 cells using Lipofectamine 2000 (Thermo Scientific, Waltham, MA, USA) by following the manufacturer’s instructions. The cells were transfected for 24 h, and then used for further tests after being treated with or without cisplatin for 24 h. The siRNA targeting Nrf2 (siNrf2) and its negative control (siNC) were obtained from RiboBio (Guangzhou, China) and transfected into cells, following the manufacturer’s instructions.
7
Knockdown of PINK1, Parkin, Nrf2, and HO-1 in Nucleus Pulposus Cells
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SiRNAs against PINK1 (siPINK1) (5′‐GCUAGUUACAAGAGAACAA‐3′), Parkin (siParkin) (5′‐GAAUACAUUCCCUACCUCA‐3′), Nrf2 (siNrf2) (5′‐GAGAAAGAAUUGCCUGUAA‐3′), and HO‐1 (siHO‐1) (5′‐CAGUUGCUGUAGGGCUUUA‐3′) and scrambled siRNA (si‐Control) (5′‐UUCUCCGAACGUGUCACGU‐3′) were designed and synthesized by RiboBio. Nucleus pulposus cells were transfected for 48 hours with 100 nmol/L of each siRNA using Lipofectamine® 2000 (Invitrogen) according to the manufacturer's instructions. The cells were then used for experiments.
8
Molecular Regulation of NRF2 and NQO1
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The expression plasmids, pENTER-NRF2, pENTER-NQO1, and the pENTER-vector, were obtained from Vigenebio Inc. (MD, USA). The siRNAs (small interfering RNAs), siNQO1, siNRF2, and siNC (negative control), and miRNAs, miR-450b-5p mimic, miR-450b-5p inhibitor, and their respective negative controls, were all purchased from Ribobio Inc. (Guangzhou, China). The plasmids, miRNAs and siRNAs, were transfected alone or co-transfected into cells as mentioned in the result part using Lipofectamine 2000 (Thermo Fisher, CA, USA) according to the manufacturer’s instructions. The sequences of siRNAs and miRNAs were listed in the Supplementary Table S2 .
9
Chondrocyte Transfection with siNrf2
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After reaching 50% confluence, chondrocytes cultured in six-well plates were transfected with siNrf2 and siNC, which were provided by RiboBio (Guangzhou, China), using Lipofectamine 3000 transfection reagent on the base of the instruction for 6 h at 37°C. Transfected cells were used for subsequent experiments.
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