KM-H2 and L-428 cells were transfected with 100 nM of either Silencer Select IER3-directed small interference RNA (siRNA) or Silencer Select Negative Control siRNA (Ambion, Carlsbad, CA, USA); SignalSilence p44/42 MAPK (ERK1/2) siRNA or Control siRNA (Cell Signaling Technology) according to the manufacturer’s instructions. Twenty-four hours after transfection, the cells were treated with 10 μM AEZS-136. The effects of gene silencing were evaluated by immunofluorescence, mRNA and cell death as described above after 24 and 48 hours of drug exposure.
Signalsilence p44 42 mapk erk1 2 sirna
Manufactured by Cell Signaling Technology
Sourced in United States
SignalSilence® p44/42 MAPK (Erk1/2) siRNA is a small interfering RNA (siRNA) product designed to target and silence the expression of the p44/42 MAPK (Erk1/2) gene. It is intended for use in cell-based assays to study the function of this gene.
Automatically generated - may contain errors
Lab products found in correlation
Control sirna, by Cell Signaling Technology (1 mentions)
Erdafitinib, by Selleck Chemicals (1 mentions)
Byl719, by Selleck Chemicals (1 mentions)
Mek inhibitor u0126, by Promega (1 mentions)
Mopc 21, by BioLegend (1 mentions)
Perm wash, by BD (1 mentions)
Pd173074, by Selleck Chemicals (1 mentions)
Cytofix cytoperm, by BD (1 mentions)
Cytoflex lx flow cytometer, by Beckman Coulter (1 mentions)
Cytexpert, by Beckman Coulter (1 mentions)
Transit x2, by Mirus Bio (1 mentions)
Sigenome smartpool egr1, by Horizon Discovery (1 mentions)
Signalsilence perk sirna 1, by Cell Signaling Technology (1 mentions)
Dharmafect 1 transfection reagent, by Horizon Discovery (1 mentions)
5 protocols using signalsilence p44 42 mapk erk1 2 sirna
1
Investigating IER3 and MAPK Modulation in Cell Lines
2
Immune Cell Profiling and Modulation
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Immune cells from mice were stained with PerCP-conjugated anti-CD4 (GK1.5, BioLegend) mAbs, APC/Cy7-conjugated anti-CD8a (53–6.7) mAbs, and the isotype monoclonal mAb. FITC-conjugated anti-I-A/I-E mAbs (M5/114.15.2, BioLegend) was used for negative gating. After pretreatment with 3 μM FGFR1-TKIs (PD173074; AZD4547; Erdafitinib, Selleck Chemicals), 3 μM mitogen-activated protein kinase (MAPK) inhibitor (MEK inhibitor U0126, Promega), MAPK siRNA (SignalSilence® p44/42 MAPK Erk1/2 siRNA, Cell Signaling Technology), 3 μM STAT3 inhibitor (S3I-201, Selleck Chemicals), or 3 µM PI3K inhibitor (BYL719, Selleck Chemicals) for 48 hr, HLA class I and HLA-DR expression on tumor cell lines was assessed via flow cytometry using anti-HLA class I antibodies (Abs) conjugated with fluorescein isothiocyanate (G46-2, BD Pharmingen) and anti-HLA-DR Abs conjugated with phycoerythrin (TU36, BD Pharmingen). HNSCC cell lines were treated with or without 50 U/ml IFN-γ for 48 hr before the assay. IgG1 (MOPC-21, BioLegend) and IgG2a (MOPC-173; BioLegend) were used as isotype controls. Intracellular IFN-γ staining were performed using Perm/WashTM (BD Pharmingen), Cytofix/CytopermTM (BD Pharmingen), APC-conjugated anti-IFN-γ mAbs (4S.B3, BioLegend), and FITC-conjugated anti-granzyme B mAbs (GB11, BioLegend). Samples were analyzed using the CytoFLEX LX flow cytometer and CytExpert (Beckman Coulter).
3
siRNA Transfection in C6 Cells
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Control siRNA, human σ-1R siRNA (sc-42250), human Src siRNA (sc-29228), human NF-κB p65 siRNA (sc-29410), and rat HMGB1 siRNA (sc-270015) were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Signal Silence® p44/42 MAPK (Erk1/2) siRNA was purchased from Cell Signaling (Danvers, MA, USA). The siRNAs were prepared according to the transfection protocol for cell cultures from Santa Cruz Biotechnology. Briefly, 1 ml of siRNA transfection reagent mixture (transfection reagent, sc-29528; transfection medium, sc-36868) was co-incubated with C6 cells for 5 h in a 5 % CO2 incubator at 37 °C, and an equal amount of DMEM with 20 % FBS was then added. An additional incubation was performed for 18 h, and the procedure for conditioned media was then performed.
4
Silencing MAPK in Rheumatoid Arthritis
5
Viral Infection Assay in Astrocytes
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Astrocytes seeded at 2.5 × 105 cells per well in a 12-well plate were transfected with 50 nM siGenome SMARTpool EGR1 (Dharmacon, Cat# M-006526-01), 100 nM SignalSilence® p44/42 MAPK (Erk1/2) siRNA (Cell Signaling, Cat# 6560S), 100 nM SignalSilence® PERK siRNA I (Cell Signaling, Cat# 9024S), or AllStar negative-control small interfering RNA (Qiagen, Cat# 1027280), using 1.2 μl of the DharmaFECT 1 transfection reagent (Dharmacon, Cat# T-2001-02). At 48 h post-transfection, cells were infected with VEEV TC-83 (MOI 5) for 1 h. After infection the medium was replaced with fresh medium. At 18hpi or 36hpi lysates were collected for analysis.
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