Human recombinant TNF-α, IL-17A and IL-1β were purchased from PeproTech (Rocky Hill, NJ, USA). Dimethyl sulfoxide (DMSO) was purchased from Nacalai Tesque (Kyoto, Japan). Sirtinol (Enzo Life Science, Farmingdale, NY, USA) and dorsomorphin (Abcam, Cambridge, UK) were dissolved in DMSO and stored at −30 °C until used in the experiments. Metformin (Metformin hydrochloride, Tokyo Chemical Industry, Tokyo, Japan) was dissolved in medium and filtrated by 0.22 µm polyvinylidene difluoride (PVDF) membrane (Merck Millipore, Burlington, MA, USA). Anti-human pro-IL-1β monoclonal rabbit antibody, anti-human NLRP3 monoclonal rabbit antibody, anti-human caspase-1 monoclonal rabbit antibody, anti-human β-actin monoclonal mouse antibody, anti-phosphorylated AMPKα rabbit monoclonal antibody (Thr172), anti-AMPKα rabbit monoclonal antibody, anti-human S100A9 monoclonal rabbit antibody (Cell Signaling Technology, Danvers, MA, USA), anti-human IL-36γ monoclonal mouse antibody, anti-human S100A7 monoclonal mouse antibody and anti-human S100A8 monoclonal rabbit antibody (Abcam) were used for western blotting.
Sirtinol
Manufactured by Enzo Life Sciences
Sourced in United States
Sirtinol is a small molecule inhibitor of the sirtuin deacetylase enzyme family. It functions by directly inhibiting the activity of sirtuin enzymes, which play important roles in cellular processes such as gene expression regulation, metabolism, and aging.
Automatically generated - may contain errors
Lab products found in correlation
Dorsomorphin, by Abcam (1 mentions)
Metformin hydrochloride, by Tokyo Chemical Industry (1 mentions)
Dimethyl sulfoxide (dmso), by Nacalai Tesque (1 mentions)
Mouse anti human β actin monoclonal antibody, by Cell Signaling Technology (1 mentions)
Il 17a, by Thermo Fisher Scientific (1 mentions)
Il 1β, by Thermo Fisher Scientific (1 mentions)
Recombinant human tnf α, by Thermo Fisher Scientific (1 mentions)
Ly294002 hydrochloride, by Bio-Techne (1 mentions)
Fk866, by Adipogen (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
3 protocols using sirtinol
1
Inflammatory Cytokine Signaling Modulation
2
Sirtinol Inhibits SIRT1 Activity
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SIRT1 activity was inhibited by treating the cells with 60 µM sirtinol (Enzo Life Science, USA)59 . Cells were initially washed with PBS and treated with sirtinol for 24 h in a 5% CO2 humidified incubator at 37 °C. Inhibition of SIRT1 at gene and protein levels was carried out in stages whereby the gene is silenced first followed by protein inhibition.
3
Metabolic Regulation in Cancer Cells
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HT29, COLO 205 and COLO 741 cells were provided by Roland Rad (TU Munich). BRAF-mutant COLO 741 were initially classified as colorectal cancer cells, but are now considered to derive from a pelvic melanoma metastasis [24] (link). Therefore, data obtained with COLO 741 are not shown except for immunoblots. HT29 cells were maintained in DMEM (high glucose), RKO cells (kindly provided by Bert Vogelstein) were maintained in MyCoy's 5A modified medium, COLO 741 and COLO 205 cells were kept in RPMI 1640 medium. All media were supplemented with 10% FCS, 1% L-glutamine and 1% penicillin/streptomycin (Invitrogen). FK866 (AdipoGen), sirtinol (Enzo Life Sciences) and the PI3K inhibitor (LY294002 hydrochloride (Tocris)) were dissolved in DMSO and stored at −20 °C. Cells were seeded at a concentration of 0.5 × 104 in 12 well plates. Thirty hours after seeding, FK866, the solvent DMSO, or FK866 combined with LY294002 were added to sub-confluent cells for 72 hours. sirtinol, the solvent DMSO, or sirtinol combined with LY294002 were added for 48 hours.
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