Cells were transiently treated with Keap1 siRNA and/or Nrf2 siRNA using Accell siRNA delivery medium (Dharmacon, USA) according to the manufacturer’s instructions. Cells (2 × 105/well) were added to wells of 12-well plates and cultivated overnight at 37 °C in an atmosphere of 5% CO2. Then, 2 µm of Keap1 and Nrf2 siRNA (Smartpool, Dharmacon) in 1× siRNA buffer (Dharmacon) were transferred to each well, cultivated for 72 h at 37 °C and 5% CO2, and lysed. Keap1 and/or Nrf2 silencing efficiency was examined by qPCR.
Accell sirna delivery medium
Manufactured by Horizon Discovery
Sourced in United States
Accell siRNA delivery medium is a proprietary formulation developed by Horizon Discovery to facilitate the efficient delivery of small interfering RNA (siRNA) into a variety of cell types. It is designed to enhance the uptake and intracellular trafficking of siRNA, enabling effective gene silencing without the need for transfection reagents or viral vectors.
Automatically generated - may contain errors
Lab products found in correlation
Acell smart pool of sirna targeting mouse, by Horizon Discovery (2 mentions)
Slc9a1 sirna, by Horizon Discovery (2 mentions)
Alpha enac scnn1a, by Horizon Discovery (2 mentions)
Sirna buffer, by Horizon Discovery (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
On targetplus smartpool sirna, by Horizon Discovery (2 mentions)
Smartpool, by Horizon Discovery (1 mentions)
B 002000 ub 100, by Horizon Discovery (1 mentions)
Mouse t cell nucleofector kit, by Lonza (1 mentions)
Amaxa nucleofector device, by Lonza (1 mentions)
Smartpool sirna, by Horizon Discovery (1 mentions)
Accell non targeting sirna, by Horizon Discovery (1 mentions)
Non targeting sirna, by Horizon Discovery (1 mentions)
Lab tekii 8 well chambered coverglass, by Thermo Fisher Scientific (1 mentions)
Cell discoverer 7 high content imaging system, by Zeiss (1 mentions)
17 protocols using accell sirna delivery medium
1
Keap1 and Nrf2 siRNA Knockdown
2
Antagomir-Mediated Silencing of miRNAs
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Antagomirs are chemically engineered single strand RNA oligonucleotides that silence specific miRNA in vitro in cultured cells and in vivo in animals efficiently [39 (link), 40 (link)]. As previously described [39 (link)], antagomirs against specific DLK1-Dio3 miRNAs were designed based on mature murine miRNA sequence from miRBase (http://www.mirbase.org/ ), and then synthesized by GE Dharmacon (Lafayette, CO, USA). The sequences of scrambled control specific DLK1-Dio3 miRNAs were listed in S1 Table .
As previously reported [40 (link)], to inhibit function of a specific miRNA in splenocytes, freshly isolated splenocytes were washed with PBS with 0.5% BSA, resuspended in serum free Accell siRNA delivery medium (GE Dharmacon) supplemented with 1μM specific antagomir or scrambled control antagomir at 10x106/ml, and then incubated in the cell incubator (37°C, 5% CO2) for 1.5–2 hrs. After incubation, the treated cells were pelleted and resuspended with complete RPMI medium supplemented with 0.1μM respective antagomirs at 5x106/ml and plated into 24 well plate for culture. Twenty-four hours after antagomirs treatment, the cells were stimulated with lipopolysaccharide (LPS, 500ng/ml, from Sigma-Aldrich) for the designated time. The supernatant were collected for analysis of cytokine production.
As previously reported [40 (link)], to inhibit function of a specific miRNA in splenocytes, freshly isolated splenocytes were washed with PBS with 0.5% BSA, resuspended in serum free Accell siRNA delivery medium (GE Dharmacon) supplemented with 1μM specific antagomir or scrambled control antagomir at 10x106/ml, and then incubated in the cell incubator (37°C, 5% CO2) for 1.5–2 hrs. After incubation, the treated cells were pelleted and resuspended with complete RPMI medium supplemented with 0.1μM respective antagomirs at 5x106/ml and plated into 24 well plate for culture. Twenty-four hours after antagomirs treatment, the cells were stimulated with lipopolysaccharide (LPS, 500ng/ml, from Sigma-Aldrich) for the designated time. The supernatant were collected for analysis of cytokine production.
3
MUC15 Silencing in Differentiated hNECs
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Differentiated hNECs were transfected with 2 μM non-targeting siRNA (NT-siRNA) or MUC15 siRNA in Accell siRNA delivery medium (GE Healthcare Dharmacon, Inc.) for 3 times at − 48, − 24 and 6 hpi. hNECs were then harvested at 24 and 48 hpi for RT-qPCR analysis.
4
Nucleofection and siRNA Silencing in OT1 T Cells
5
Splenic DCs siRNA Knockdown Assay
6
Efficient siRNA and shRNA Knockdown of VPS35
7
Silencing Polκ in DRG Neurons
8
Targeted Silencing of mNCX-1 in Neurons
9
Genetic Manipulations in Cell Lines
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Retrovirus-mediated and lentivirus-mediated gene transductions were performed as described previously40 (link). To generate HT-LC3 U-2 OS and HeLa cells, cells were transduced with lentiviruses encoding HT-LC3 and selected with 1 µg/ml puromycin for 5 days. ATG2A/B double-knockout and ATG7 knockout U-2 OS cells were generated as previously described39 (link). To generate ATG13 knockout U-2 OS cells, cells were transfected with an equal amount (6 µg each per a 10-cm dish) of three human ATG13 gRNA for 48 h and sorted for GFP-positive transfected cells. Fourteen days after transfection, the cells were re-sorted for GFP-negative population to eliminate Cas9 stable transfectants and used for experiments. For siRNA screening, cells were grown overnight on the Lab-TekII 8-well Chambered Coverglass (Nunc, 155409) and incubated in Accell siRNA Delivery Medium (Dharmacon, B-005000-100) containing 1 µM Accell siRNA for 72 h. siRNA-mediated gene silencing was performed by nucleofection according to the manufacture’s protocol.
10
Macrophage cAMP Signaling Modulation
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Human monocyte–derived macrophages were seeded onto 12-well plates at 5 × 105 cells per well and then transfected with 1 μM Accell Human Control siRNA or Accell Anti–human GPR101 siRNA SMARTpool for 72 hours in serum-free Accell siRNA Delivery Medium (Dharmacon). Cells were incubated with RvD5n-3DPA (1 nM) or vehicle (PBS containing 0.01% ethanol) for 2 minutes, after which incubations were quenched with 5% Triton X-100, cells were homogenized, and cAMP levels were determined by ELISA following the manufacturer’s instruction (Elite cAMP ELISA Assay kit, eEnzyme).
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