Schwann cells were transfected with siRNA using Lipofectamine™ RNAi MAX complexes (Invitrogen) at 4 or 5 days post-purification. ON-TARGET plus Non-targeting control siRNA (catalog D-0018100-01-20, GE Dharmacon, Lafayette, CO, USA) served as the control. The siRNA primer sequences for Rab27a and siRNA Slp2-a are listed in Table 2
Lipofectamine rnai max complexes
Manufactured by Thermo Fisher Scientific
Lipofectamine™ RNAi MAX complexes are a transfection reagent designed for efficient delivery of small interfering RNA (siRNA) or short hairpin RNA (shRNA) into mammalian cells. The complexes facilitate the introduction of RNA interference (RNAi) molecules into the cells, enabling the study of gene function through gene silencing.
Automatically generated - may contain errors
Lab products found in correlation
On targetplus non targeting control sirna, by Horizon Discovery (1 mentions)
On targetplus nontargeting control pool, by GE Healthcare (1 mentions)
On targetplus smartpool, by GE Healthcare (1 mentions)
Tas1r3 sirna, by Bioneer (1 mentions)
Dulbecco s modified eagle s medium f 12, by Welgene (1 mentions)
3 protocols using lipofectamine rnai max complexes
1
Knockdown of Rab27a and Slp2-a in Schwann Cells
2
Silencing IRF1 and IRF3 in A549 Cells
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siRNAs for human IRF1 (3659, ON-TARGETplus smart pool including 4 target sequences: GGGCUCAUCUGGAUUAAUA, UGAACUCCCUGCCAGAUAU, GCUCAGCUGUGCGAGUGUA, GAAGGGAAAUUACCUGAGG), siRNA for human IRF3 (3661, ON-TARGETplus smart pool including 4 target sequences: CGAGGCCACUGGUGCAUAU, CCAGACACCUCUCCGGACA, GGAGUGAUGAGCUACGUGA, AGACAUUCUGGAUGAGUUA) and ON-TARGETplus non-targeting control pool were purchased from GE health, Dharmacon. 3 x 104 A549 cells were transfected with 25 μM of different siRNAs with Lipofectamine RNAiMAX complexes (Invitrogen) according to the manufacture’s instruction (reverse transfection). After 16 h of incubation, media was replaced by complete cell culture media without antibiotics. After 40 h of transfection, cells were infected with RSV-HD at a moi of 1.5 TCID50/cell for 10 h. Cells were harvest with either TRIzol for RNA or NP-40 lysis buffer for protein analysis. As control, cells were treated only with Lipofectamine RNAiMAX transfection reagent.
3
Silencing Tas1r3 in Murine TM3 Cells
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Murine TM3 cells (Cat# CRL-1714; American Type Culture Collection, Manassas, VA, USA) were cultured at 37 °C with 5 % CO2 in Dulbecco's modified Eagle's medium/F-12 (Cat# LM002-04; WelGENE Inc., Daegu, Korea), with 10 % fetal bovine serum and 1 % penicillin-streptomycin solution added. For knocking down Tas1r3, 1 × 105 TM3 cells per well were seeded in a 6-well culture plate approximately 24 h before transfection. Upon reaching 60 % confluency, each well was transfected with Tas1r3 siRNA (20 nM; Bioneer, Daejeon, South Korea) and Lipofectamine™ RNAiMAX® complexes (Invitrogen, Thermo Fisher Scientific, USA) according to the manufacturer's instructions. Following 48 h incubation, all media were collected, and the cells were harvested for subsequent biochemical analysis. The siRNAs' target sequences utilized for the knockdown are detailed in Table S2 .
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