Transient transfection was performed using the Lipofectamine Reagent (RNAiMAX, Life Technologies) as per manufacturer's instructions. In short, THP1 cells with PMA added were seeded at approximately 80% confluency. The next day silencing RNA molecules (siRNA) were used at a final concentration of 20nM to transfect the cells. Cells were cultured for 24 hours in the presence of the transfection reagents prior to infection and LPS treatment, the following, pre-validated siRNAs used were: IRAK-M siRNA (sc-39098, Santa-Cruz), PPARγ siRNA (sc-29455, Santa-Cruz), CD26 siRNA (sc-42762, Santa-Cruz) and Silencer Negative Control (SCR) siRNA (AM4611g, Ambion).
Pparγ sirna
Manufactured by Santa Cruz Biotechnology
Sourced in United States
PPARγ siRNA is a laboratory reagent used for gene silencing experiments. It is designed to target and downregulate the expression of the PPARγ (Peroxisome Proliferator-Activated Receptor Gamma) gene. This tool can be utilized in various cell-based research applications to investigate the functional role of the PPARγ gene.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine reagent, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Pparγ, by Thermo Fisher Scientific (1 mentions)
Oligofectamine reagent, by Thermo Fisher Scientific (1 mentions)
Egr1 sirna, by Santa Cruz Biotechnology (1 mentions)
Ampkα, by Santa Cruz Biotechnology (1 mentions)
Control nonspecific sirna oligonucleotides, by Santa Cruz Biotechnology (1 mentions)
Jagged 1 sirna, by Santa Cruz Biotechnology (1 mentions)
β catenin sirna, by Santa Cruz Biotechnology (1 mentions)
5 protocols using pparγ sirna
1
Transient Transfection of THP1 Cells
2
Silencing Akt and PPARγ Pathways in HUVECs
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SignalSilence Akt siRNA (Cell Signaling Technology, Inc. Danvers, MD, USA) or PPARγ siRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were transfected into HUVECs using Lipofectamine according to the protocol. Cells (40% confluent) were serum-starved for 1 h followed by incubation with 100 nM target siRNA or control siRNA for 6 h in serum-free media. The serum-containing media was then added (10% serum final concentration) for 42 h before experiments and/or functional assays were conducted. Target protein silencing was assessed through protein analysis up to 48 h after transfection.
3
PPARγ Knockdown in HepG2 Cells
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Transient transfection of HepG2 cells was performed using siRNAs. Briefly, 24 h after plating, cells were transfected with PPARγ siRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) or an siRNA scrambled control (Santa Cruz Biotechnology, Santa Cruz, CA, USA) (50 or 100 nM) using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA). At 48 h after transfection, the levels of PPARγ mRNA/protein were analyzed by quantitative RT-PCR and Western blotting.
4
Silencing PDPK1, Egr-1, and PPARγ in Cell Signaling
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The siRNA human PDPK1 (EHU071261) was ordered from Sigma (Shanghai, China). The AMPKα (Cat No.sc-45312), Egr-1 siRNA (Cat No. sc-105070), PPARγ siRNA (Cat No. sc-29455), and control nonspecific siRNA oligonucleotides (Cat No. sc-37007) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). For the transfection procedure, cells were grown to 60% confluence, and PDK1, Egr-1, and PPARγ and control siRNAs were transfected using the oligofectamine reagent (Invitrogen, Shanghai, China) according to the manufacturer’s instructions. Briefly, Lipofectamine was incubated with serum–free medium for 10 min., mixed with siRNA (80 nM), incubated for 20 min at room temperature before the mixture was diluted with medium and added to cells. After culturing for 30 h, cells were washed, resuspended in new culture media in the presence or absence of ciglitazone for an additional 24 h for Western Blot, cell growth, luciferase report assays and other experiments.
5
Murine Bone Marrow-Derived Macrophage Transfection
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