Nonspecific, β‐catenin, metadherin, and p65 siRNAs were purchased from Dharmacon (Lafayette, CO). Cells were transfected with 20 nmol L‐1 siRNA using siLectFect (Bio‐Rad, Hercules, CA) as instructed by the manufacturer.
Silectfect
Manufactured by Bio-Rad
Sourced in United States
SiLectFect is a cationic lipid-based transfection reagent for the delivery of small interfering RNA (siRNA) and other nucleic acids into mammalian cells. It facilitates the efficient uptake and transfection of these molecules, enabling gene silencing and other applications.
Automatically generated - may contain errors
4 protocols using silectfect
1
Targeted Gene Silencing Via siRNA
2
Targeted Gene Silencing in Cell Lines
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Cells were seeded in six-well plates 24 h before transfection. 20 nM Dicer siRNA (Dharmacon, Catalog No. D-003483), Drosha siRNA (Dharmacon, Catalog No. D-016996), Exportin-5 siRNA (Dharmacon, Catalog No. D-014000), Akt siRNA (Dharmacon, Catalog No. D-003000), or c-Met siRNA (Dharmacon, Catalog No. L-003156) was transfected with 1.5 μl siLectFect (Bio-Rad) for 24 h. Similar protocol was used for transfecting miR-199a-3p mimic (Ambion, Catalog No. 4464066) at 10 nM concentration. Cells were at 80% confluence at the time of transfection. Respective non-specific controls were incorporated in each experimental setup.
3
Silencing Nur77, β-catenin, and HIF-1α
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Small interfering RNA (siRNA) targeting Nur77 (SMARTpool 5′-UCGAGGACUUCCAGGUGUA-3′, 5′-GGACAGAGCAGCUGCCCAA-3′, 5′-GAAGGCCGCUGUGCUGUGU-3′, 5′-CGGCUACACAGGAGAGUUU-3′), β-catenin (5′-AAGUCCUGUAUGAGUGGGAAC-3′), HIF-1α (5′-GGACACAGAUUUAGACUUG-3′), and a non-specific duplex oligo (5′-GGCTACGTCCAGGAGCGCA-3′) were purchased from Dharmacon (Lafayette, CO, USA). Cells were transfected with 20 nmol l−1 siRNA using siLectFect (Bio-Rad, Hercules, CA, USA) following the manufacturer's instructions. Transfected cells were serum starved for 24 h before incubation under hypoxic conditions.
4
Nur77, Dicer, p63 siRNA Transfection
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Small interfering RNA (siRNA) targeting Nur77 (SMARTpool), Dicer (SMARTpool), p63 (SMARTpool), and an NS duplex oligo (5'-GGC TAC GTC CAG GAG CGC A-3') were purchased from Dharmacon (Lafayette, CO, USA). Let-7i-5p, non-specific miRNA mimic and let-7i-5p inhibitor were purchased from Ambion (Austin, TX, USA). Cells were transfected with 20 nM siRNA, miRNA mimic or miRNA inhibitor using siLectFect (Bio-Rad, Hercules, CA, USA) following the manufacturer's instructions.
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