Bone marrow cells were differentiated in the presence of recombinant mouse M-CSF (20 ng/ml; R&D Systems) in complete medium (RPMI 1640 containing 10 mM glucose, 2 mM L-glutamine, 100 U/mL of penicillin/streptomycin and 10% FBS) for 7 days. Day 7 macrophages were washed and variously stimulated with IL-4 (20 ng/ml; PeproTech) or lipopolysaccharide (LPS, 20 ng/ml; Sigma) plus IFN-γ (50 ng/ml; R&D Systems) in the presence or absence of 200 μM etomoxir (ETO; Sigma), 20 μM sulfo-n-succinimidyl oleate (SSO), 30 μM chloroquine (CLQ; Sigma), 0.1 μM bafilomycin A1 (Baf; Tocris Bioscience), 100 μM Orlistat (Cayman), 20 μM 5-(Tetradecyloxy)-2-furoic Acid (TOFA, Sigma), 5 or 50 μg/ml of LDL (Kalen Biomedical, LLC) and VLDL (Kalen Biomedical, LLC) for 24 h. Macrophages were then harvested and analyzed by flow cytometry for expression of markers of M1 or M2 activation. In some experiments the survival of macrophages was monitored following M2 or M1 activation. For these experiments, bone marrow-derived macrophages (0.5 × 106 cells per well of a 48-well plate) were cultured in complete medium with mouse M-CSF (20 ng/ml) and stimulated with IL-4 or LPS plus IFN-γ. Cell viability was monitored by flow cytometric analysis of 7-amino-actinomycin D (7-AAD; BioLegend) staining of F4/80+ cells over time. Culture medium and stimulation signals were replenished every 1 or 2 days.
5 tetradecyloxy 2 furoic acid tofa
Manufactured by Merck Group
Sourced in United States
5-(Tetradecyloxy)-2-furoic Acid (TOFA) is a chemical compound used as a laboratory reagent. It is a colorless solid with a faint odor. TOFA has a molecular formula of C_{19}H_{34}O_{3} and a molecular weight of 326.47 g/mol.
Automatically generated - may contain errors
Lab products found in correlation
7 aminoactinomycin d 7 aad, by BioLegend (2 mentions)
Chloroquine clq, by Merck Group (2 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (2 mentions)
Sulfo n succinimidyl oleate, by Merck Group (2 mentions)
Bafilomycin a1 baf, by Bio-Techne (2 mentions)
Recombinant mouse m csf, by R&D Systems (2 mentions)
Orlistat, by Cayman Chemical (2 mentions)
Etomoxir eto, by Merck Group (2 mentions)
Ifn γ, by R&D Systems (2 mentions)
Gemfibrozil, by Santa Cruz Biotechnology (1 mentions)
Cpg a, by InvivoGen (1 mentions)
Cytometric bead array cba, by BD (1 mentions)
Mar1 5a3, by Leinco Technologies (1 mentions)
Mouse ifn α elisa kit, by PBL Assay Science (1 mentions)
Ifn α, by PBL Assay Science (1 mentions)
Uk5099, by Merck Group (1 mentions)
Poly 1 c, by Merck Group (1 mentions)
Simvastatin, by Merck Group (1 mentions)
Cpi 613, by Selleck Chemicals (1 mentions)
P ulk1, by Cell Signaling Technology (1 mentions)
7 protocols using 5 tetradecyloxy 2 furoic acid tofa
1
Macrophage Polarization and Survival Assay
2
Modulating dendritic cell responses
3
Macrophage Polarization and Survival Assay
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Bone marrow cells were differentiated in the presence of recombinant mouse M-CSF (20 ng/ml; R&D Systems) in complete medium (RPMI 1640 containing 10 mM glucose, 2 mM L-glutamine, 100 U/mL of penicillin/streptomycin and 10% FBS) for 7 days. Day 7 macrophages were washed and variously stimulated with IL-4 (20 ng/ml; PeproTech) or lipopolysaccharide (LPS, 20 ng/ml; Sigma) plus IFN-γ (50 ng/ml; R&D Systems) in the presence or absence of 200 μM etomoxir (ETO; Sigma), 20 μM sulfo-n-succinimidyl oleate (SSO), 30 μM chloroquine (CLQ; Sigma), 0.1 μM bafilomycin A1 (Baf; Tocris Bioscience), 100 μM Orlistat (Cayman), 20 μM 5-(Tetradecyloxy)-2-furoic Acid (TOFA, Sigma), 5 or 50 μg/ml of LDL (Kalen Biomedical, LLC) and VLDL (Kalen Biomedical, LLC) for 24 h. Macrophages were then harvested and analyzed by flow cytometry for expression of markers of M1 or M2 activation. In some experiments the survival of macrophages was monitored following M2 or M1 activation. For these experiments, bone marrow-derived macrophages (0.5 × 106 cells per well of a 48-well plate) were cultured in complete medium with mouse M-CSF (20 ng/ml) and stimulated with IL-4 or LPS plus IFN-γ. Cell viability was monitored by flow cytometric analysis of 7-amino-actinomycin D (7-AAD; BioLegend) staining of F4/80+ cells over time. Culture medium and stimulation signals were replenished every 1 or 2 days.
4
Metabolic Modulation in Cancer Cells
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CPI-613 was obtained from Selleckchem (Houston, TX). 2′,7′-dichlorofluorescin diacetate (DCFH-DA), chloroquine (CQ), N-acetylcysteine (NAC), Compound C, simvastatin, and 5-(tetradecyloxy)-2-furoic acid (TOFA) were purchased from Sigma-Aldrich (St Louis, MO). Fatty acid and lipid metabolism antibody sampler kit and glycolysis antibody sampler kit were purchased from Cell Signaling Technology (Beverly, MA). Fatty acid and lipid metabolism antibody sampler kit includes antibodies against Acetyl-CoA Carboxylase (ACC), p-Acetyl-CoA Carboxylase (p-ACC) (Ser79), AceCS1, ACSL1, Lipin 1, ATP-Citrate Lyase, p-ATP-Citrate Lyase, Fatty Acid Synthase (FAS). glycolysis antibody sampler kit includes GAPDH, PDH, HKI, HKII, LDHA, PKM2, PKM1/2, and PFKP. Antibodies against c-Caspase 3, PARP, p-mTOR (Ser2448), mTOR, p62, LC3B, p-AMPKα (Thr172), AMPKα, p-ULK1, ULK1, Bax and Bcl-2 were purchased from Cell Signaling. Apoptotic rate was determined using Annexin V-FITC Apoptosis Detection Kit (BD Biosciences, San Jose, CA). All flow cytometry data was analyzed using FlowJo software (Tree Star, Ashland, OR). Cell viability was determined by MTT assay, crystal violet staining, and alamarBlue Cell Viability Reagent (Thermo Fisher Scientific, Waltham, MA). Western blot, plasmid transfection and lentiviral infection were carried out as we previously described [15 (link)–17 ].
5
Metabolic Modulation of T Cell Functionality
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Memory CD4+ T cells were cultured in complete RPMI (RPMI (Labtech) supplemented with 10% FCS (Sigma Aldrich), 2 mM L-glutamine with 1% penicillin/streptomycin (Sigma Aldrich) with irradiated antigen-presenting cells (irrAPC), anti-CD3 (eBioscience) and, where indicated, in the presence of dichloroacetate (DCA) (10 mM, Sigma Aldrich), rapamycin (20 nM, Sigma Aldrich), 5-(Tetradecyloxy)-2-furoic acid (TOFA) (1.2 μg/ml, Sigma Aldrich), C-75 (0.6 μg/ml, Sigma Aldrich), cerulenin (3.1 μM, Sigma Aldrich), or palmitate (25 μM, Seahorse Biosciences) for 5 days. Isolated CD161+, CD161−, and Tconv cells were cultured with irradiated antigen presenting cells (irrAPC), anti-CD3 and IL-2 (20 u/ml, eBioscience) for 6 days or in some cases with PMA/ionomycin for 18 h prior to Seahorse analysis. Isolated Treg cells were cultured using the Treg cell expansion kit (Miltenyi Biotec) in the presence of IL-2 (500 u/ml) for 6 days prior to Seahorse analysis. During glucose deprivation, memory CD4+ T cells were cultured in glucose-free complete RPMI (RPMI (Biosciences) supplemented with 10% dialysed FCS (Sigma Aldrich), 1% vitamin cocktail (InvivoGen), and 1% selenium/insulin cocktail (InvivoGen) supplemented with either 10 mM glucose (Sigma Aldrich) or 10 mM galactose (Sigma Aldrich) for 5 days.
6
Peptide Synthesis and Pheromone Quantification
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The PBAN-like peptide (Ser-Arg-Thr-Lys-Tyr-Phe-Ser-Pro-Arg-Leu-NH2) was synthesized by Sangon Biotech Company (Shanghai, China). O. furnacalis sex pheromone components, (E)-tetradec-12-enyl acetate (E12-14: OAC) and (Z)-tetradec-12-enyl acetate (Z12-14: OAC), were purchased from Sigma Company (St. Louis, MO, USA) and were used to quantify sex pheromone titer in the PGs by gas chromatography (GC). The 5-(tetradecyloxy)-2-furoic acid (TOFA, an ACC inhibitor), LaCl3 (a Ca2+ inhibitor), and tacrolimus (FK506, CaN-specific inhibitor) were purchased from Sigma (St. Louis, MO, USA).
7
Endoplasmic Reticulum Stress Pathway
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ALA, PA, DMSO, and 5-tetradecyloxy-2-furoic acid (TOFA) were purchased from Sigma (St. Louis, MO, USA). RPMI 1640 Medium basic and phosphate-buffered saline (PBS) were purchased from Biological Industries (Israel). Fetal bovine serum (FBS) was purchased from South America sterile filtered. Antibodies of FASN, IRE1, ATF6, PERK, and CHOP were purchased from Cell Signaling Technology (Denvers, MA, USA).
GAPDH was purchased from Lablead (Beijing, China).
GAPDH was purchased from Lablead (Beijing, China).
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