SiRNA transfection assay was performed by the N-TERTM Nanoparticle siRNA Transfection System following the manufacturer's protocol (Sigma-Aldrich, St. Louis, MO, USA). MDA-MB-231 cells were transfected with siRNA Ctrl or mixture of three IL20RA siRNAs, the final concentration of siRNA used was 25 nM. After 48 h of transfection, the cells were harvested and then subjected to the follow-up experiments. The siRNA sequences were summarized in Table S4
N tertm nanoparticle sirna transfection system
Manufactured by Merck Group
Sourced in United States
The N-TERTM Nanoparticle siRNA Transfection System is a laboratory tool designed for the delivery of small interfering RNA (siRNA) into cells. It utilizes nanoparticles to facilitate the efficient introduction of siRNA into target cells, enabling the study of gene function and regulation.
Automatically generated - may contain errors
4 protocols using n tertm nanoparticle sirna transfection system
1
siRNA Transfection Assay for MDA-MB-231 Cells
2
Silencing α-Taxilin in Huh7.5 Cells
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Hepatitis C virus-negative Huh7.5 cells were transfected 4 h after seeding either with 50 nM α-taxilin specific siRNA or scrRNA (sc-39644 and sc-37007, Santa Cruz Biotechnology) using N-TERTM Nanoparticle siRNA Transfection System (Sigma) according to the manufacturer’s protocol. For controls, cells were again transfected after 24 h of transient RNAi transfection with empty plasmid pUC18 or pDest26-TXLNA using PEI (polyethyleneimine; Polyscience). Cells were harvested three days post-transfection with siRNA.
3
Targeted Silencing of ERK1/2 in Cell Cultures
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Control, ERK1, and ERK2 small interfering RNAs (siRNA) were purchased from Santa Cruz
Biotechnology (USA). Transfections were performed with the N-TERTM Nanoparticle siRNA
Transfection System (Sigma). Briefly, 1×106 cells were seeded into each
well of 6-well plates and cultured for 48 h. Transfection was carried out for 12 h
after adding a nanoparticle formation solution containing 20 nM target siRNA to each
well. The cells were then maintained in conventional cultures for 24 h before
conducting further experiments.
Biotechnology (USA). Transfections were performed with the N-TERTM Nanoparticle siRNA
Transfection System (Sigma). Briefly, 1×106 cells were seeded into each
well of 6-well plates and cultured for 48 h. Transfection was carried out for 12 h
after adding a nanoparticle formation solution containing 20 nM target siRNA to each
well. The cells were then maintained in conventional cultures for 24 h before
conducting further experiments.
4
siRNA Transfection in Oocytes
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DNA templates for siRNA and siRNA production are described in Supplementary File 1 . For siRNA transfection, the N-TERTM Nanoparticle siRNA Transfection System (Sigma) was used. Briefly, two tubes, one containing 1.1 μl N-TERTM nanoparticles in 5.15 μl nuclease-free water (Acros Organics) and the other containing 1.625 μl of siRNA (5 μM) mixture in 4.625 μl of siRNA dilution buffer (provided by the kit) were gently mixed together and incubated at room temperature (RT) for 20 min. Next, the siRNA–nanoparticle complex solution was added to 100 μl of medium containing 50 oocytes. After 12–14 h, the oocytes were washed to remove the nanoparticle-containing medium. After 1–2 h, siRNA treatment was repeated, depending on how difficult the target was knocked down. 2.5 nM milrinone was added to cultures during siRNA treatment to prevent resumption of meiosis. Next, oocytes were transferred into milrinone-free MEM + and cultured for 8 or 16 h.
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