RAW264.7 cells were cultured to 50%–60% confluence in DMEM without penicillin/streptomycin. The cells were then transfected with 100 nmol/L Nlrp3 siRNA (sc-45470; Santa Cruz Biotechnology Inc., Dallas, TX, USA) or control nonspecific siRNA (sc-37007; Santa Cruz Biotechnology Inc.) using Lipofectamine 2000 (Thermo Fisher Scientific) according to the manufacturer’s instructions. After transfection for 24 hours, the cells were exposed to 2 µg/cm2 for 12 hours.
Nlrp3 sirna
Manufactured by Santa Cruz Biotechnology
Sourced in United States
NLRP3 siRNA is a small interfering RNA designed to target the NLRP3 gene, which encodes a protein involved in the formation of the NLRP3 inflammasome. The core function of this product is to facilitate the specific knockdown of NLRP3 gene expression in experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Control sirna, by Santa Cruz Biotechnology (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Sc 37007, by Santa Cruz Biotechnology (1 mentions)
Scramble sirna, by Santa Cruz Biotechnology (1 mentions)
Lipofectamine, by Thermo Fisher Scientific (1 mentions)
Gapdh sirna, by Thermo Fisher Scientific (1 mentions)
Block it alexa fluor red fluorescent control, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (1 mentions)
6 protocols using nlrp3 sirna
1
Nlrp3 Silencing in RAW264.7 Cells
2
Investigating NLRP3 Silencing in Melanoma Cells
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For siRNA transfection, 1205Lu, A357, and HS294T cells (2 × 105) were incubated with scrambled or NLRP3 siRNA (Santa Cruz Biotechnology). Transfection of the siRNA duplexes (2 nM) was carried out using siRNA Transfection Medium according to the manufacturer’s instructions. After 24 h, the medium was replaced with RPMI 10% FBS (500 µL), and the cells were incubated for an additional 24 h. The supernatants were collected for the measurement of IL-1β levels. Efficacy of the NLRP3 silencing was determined by Western blotting in the cell lysates.
3
NLRP3 Silencing in Monocytes
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Monocytes were transfected with either NLRP3 siRNA or control siRNA (Santa Cruz Biotechnology, Inc.) for 48 hours according to the manufacturer protocol. Briefly, 0.5 μg of siRNA was mixed with 5 μl of transfection reagent in final volume of 1 ml of transfection medium. Monocytes were incubated in transfection medium for 4 hours, washed and incubated in culture medium for 48 hours prior to infection.
4
Silencing NLRP3 in HK-2 Cells
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HK-2 cells were maintained in DMEM/F12 medium, supplemented with 10% fetal bovine serum at 37°C and 5% CO2 in a humidified incubator. Transient transfection of HK-2 cells with siRNA was previously demonstrated [17 (link)]. HK-2 cells were grown to 60% confluence and then transfected with 500 nM NLRP3 siRNA or scramble siRNA (Santa Cruz, CA) using lipofectamine (Invitrogen, CA), prior to Aldo treatment.
5
Investigating TRPM2, NLRP3, and Oxidative Stress
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Cells were transiently transfected with TRPM2 specific small interfering RNA (siRNA) (60 nmol/L; Ambion, US), NLRP3 siRNA (100 nmol/L; Santa Cruz Biotechnology, US), ASC siRNA (100 nmol/L; Ambion, US), caspase-1 siRNA (100 nmol/L; Ambion, US) or p47 phox siRNA (80 nmol/L; Ambion, US) by using Lipofectamine® RNAiMAX Transfection reagent (Gibco, US). The protocol was synthesized according to the manufacturer’s protocol. GAPDH siRNA was used as a control (40 nmol/L; Ambion, US). Transfection efficiency was > 70% assessed by BLOCK-iT™ Alexa Fluor® Red Fluorescent Control (Ambion, US) and western blotting. Cells were transfected with siRNA for 24 h before experiments.
6
NLRP3 Knockdown in HSC-T6 Cells
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HSC-T6 cells were seeded into a 6-well plate at a density of 2 × 105, and transfected with NLRP3 siRNA and control siRNA (Santa Cruz, Texas, America), according to the manufacturer’s protocol. In brief, Solution A and Solution B were prepared, mixed and incubated for 30 minutes in room temperature. Solution A: 1 µg siRNA duplex was added into 100 µl siRNA transfection medium. Solution B: 8 µl transfection reagent was added into 100 µl siRNA transfection medium.
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