Recombinant mouse mTNFα (T) (Cell sciences Cat# CRT192C) was resuspended in PBS at a concentration of 100 μg/ml. zVAD.fmk (Z) (Sigma, Cat# V116) was dissolved in DMSO at a concentration of 20 mM and used at a final concentration of 20 μM. Cycloheximide (Sigma, Cat# C-6255) was resuspended in DMSO at a concentration of 10 mg/ml and used as indicated. 7-Cl-O-Nec-1 (Nec-1s) was made by custom synthesis, dissolved in DMSO at a concentration of 10 mM and used at a final concentration of 10 μM. Custom-synthesized (Selleckchem) Smac mimetic SM-164 54 was resuspended in DMSO at a concentration of 10 mM and used at a final concentration of 500 nM. 5Z-7-Oxozeaenol (Sigma, Cat# 09890) was dissolved in DMSO at a concentration of 25 mM and used at a final concentration of 500 nM.
5z 7 oxozeaenol
Manufactured by Merck Group
Sourced in United States
5Z-7-oxozeaenol is a chemical compound that inhibits the enzymatic activity of transforming growth factor-beta-activated kinase 1 (TAK1). It is commonly used as a research tool in various scientific studies to investigate cellular signaling pathways and biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Cycloheximide (chx), by Merck Group (7 mentions)
Sp600125, by Cell Signaling Technology (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Recombinant mouse mtnfα t, by Cell Sciences (2 mentions)
Zvad fmk z, by Merck Group (2 mentions)
Custom synthesized, by Selleck Chemicals (2 mentions)
Smac mimetic sm 164 54, by Selleck Chemicals (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Y 27632, by Merck Group (2 mentions)
Necrostatin 1, by Merck Group (2 mentions)
Pd98059, by Cell Signaling Technology (2 mentions)
Celltracker green cmfda dye, by Thermo Fisher Scientific (2 mentions)
Lcl161 smac mimetic, by Selleck Chemicals (2 mentions)
Gsk 872, by Bio-Techne (2 mentions)
Z vad fmk, by Enzo Life Sciences (2 mentions)
Oligomycin, by Merck Group (2 mentions)
Pf 4708671, by Merck Group (2 mentions)
Akts473, by Cell Signaling Technology (2 mentions)
S6k1t389, by Cell Signaling Technology (2 mentions)
44 protocols using 5z 7 oxozeaenol
1
Inducing Apoptosis in Cells via Modulators
2
Alveolar Macrophage and Fibroblast Culture
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Rat alveolar macrophage NR8383 cells (CRL-2192, ATCC, Manassas, VA, USA) were cultured at 37 °C in a humidified environment containing 5% CO2 in Ham’s F-12K (Kaighn’s) medium (Life Technologies, Grand Island, NY, USA), supplemented with 15% fetal bovine serum (FBS) (Life Technologies). Human lung fibroblast WI-38 cells (ATCC; no. CCL-75) were cultured at 37 °C in a humidified environment containing 5% CO2 in Eagle's minimum essential medium (ATCC) supplemented with 10% FBS and 100 U ml–1 penicillin–streptomycin (Life Technologies). Resveratrol with a purity ⩾99% (high-performance liquid chromatography (HPLC)) and 5Z-7-oxozeaenol with a purity ⩾98% (HPLC) were purchased from Sigma (St Louis, MO, USA). Chalcone derivative (4′-hydroxychalcone) with a purity ⩾98% (HPLC) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Silica particles, silicon dioxide (SiO2), with an average diameter of 1.6 μm were purchased from US Silica MIN-U-SIL-5, US Silica, Shanghai, China. Human recombinant TGF-β (TGF-β1) was purchased from Merck Millipore (Danvers, MA, USA). Cross-species TAK1 siRNA and NC siRNA were purchased from Cell Signaling Technology (Danvers, MA, USA) and transfected into cells using the Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), according to the manufacturer’s instructions. SIRT1 Activity Assay Kit was from Abcam (Cambridge, MA, USA).
3
Inhibition of TAK1 Pathway
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The TAK1 inhibitor 5Z-7-oxozeaenol (Sigma) was diluted in dimethyl sulfoxide (DMSO) and used at a concentration of 300 nM. Puromycin and hygromycin (Sigma) were used at 5 μg/ml and 50 μg/ml, respectively. Mouse anti-c-Myc and anti-β-actin antibodies were from Sigma. Rabbit anti-TAK1 and anti-β-tubulin antibodies were from Cell Signaling Technologies.
4
Intracerebroventricular Delivery of TAK1 Inhibitor
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The specific TAK1 inhibitor (5Z‐7‐oxozeaenol; Sigma‐Aldrich, O9890) was dissolved in dimethyl sulfoxide (DMSO, Sigma‐Aldrich, D2650) (0.8 μg/μL), as previously described.25 2 μL of 5Z‐7‐oxozeaenol solution was administered into the intracerebroventricular of non‐transgenic and CARD3‐TG mice 30 minutes before tMCAO through stereotaxic apparatus (Stoelting, Wood Dale, IL, 51900). An equal volume of DMSO was administered as control treatment.
5
Detailed Antibody-Based Immunoassay Protocol
6
Modulation of A431 Cell Signaling
7
Modulation of Cell Death Pathways
8
Influenza A Virus Infection in MEFs
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Murine embryonic fibroblasts (MEFs) were obtained from 13- to 14-days-old WT mouse embryos digested with 0.5% trypsin-EDTA solution for 30 minutes at 37°C [24 (link)]. MEFs were cultured in MEM supplemented with 20% heat inactivated FBS. Cells were pretreated for 30 minutes with specific TAK1 inhibitor 5Z-7-oxozeaenol (1 μM) (Sigma-Aldrich, Oakville, ON, Canada), infected with Influenza A virus at 0.5 multiplicity of infection (m.o.i.) and treated with placebo or LTB4 (1 μM). Cells were also stimulated with MDP alone (10 μg/ml) or in combination with LTB4 (1 μM). Supernatants and cells were collected 6 hours post-treatment. Protein samples were extracted from cells and assayed for western blot analyses, as detailed above, using anti-IPS–1, anti-phospho-IRF3, anti-phospho-IRF7 and anti-NF-κB p65 antibodies. Supernatants were assayed for IFNβ, TNFα and IL–6 by ELISA.
9
Disrupting IRF3 and NF-kB Signaling
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Pharmacological synthetic inhibitors were used to block the proteins that regulate interferon regulatory factors (IRFs) and NF-kB downstream signalling pathways. The TBK1-mediated activation of IRF3 was blocked by 0.1 μM BX795 [30 (link)], IkB kinase (IKK)-mediated NF-kB activation by 10 nM PS1145 [31 (link)] and TAK1 activation by 0.1 μM 5Z-7-oxozeaenol [32 (link)] (Sigma-Aldrich, Stockholm, Sweden). The cells were incubated at 37°C for 1 h in the presence of all inhibitors and then infected with 1 MOI RV1B for 24 h.
10
Antibody Panel for ADAM12 and Signaling
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The following antibodies were employed in this study: mouse ADAM12 (Proteintech 14139-1-AP); human ADAM12 (Sigma HPA030867); human TAK1 (SCBT sc-1839); human KAT2A (SCBT sc-20698); human SIRT6 (Abcam ab62739); human SMAD3 (ab28379), human phospho-SMAD3 (Abcam ab52903), human tubulin (Abcam ab7291), human TAB1 (CST 3226); human Histone H3 (CST 2650). TGF-β was from Proteintech and the TAK1 inhibitor (5Z)-7-oxozeaenol from Sigma.
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