Thyroxine, KI, NaI, Lugol’s iodine (potassium triiodide), indomethacin, retinol, 9-cis-retinoic acid, and dimethyl sulfoxide (DMSO) were from Sigma. H2O2 was purchased from the local pharmacy.
Thyroxine
Manufactured by Merck Group
Sourced in United States
Thyroxine is a laboratory product used for the measurement of thyroid hormone levels in biological samples. It is a key component in the analysis of thyroid function and is widely used in clinical diagnostics and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Insulin, by Merck Group (5 mentions)
Progesterone, by Merck Group (4 mentions)
Dexamethasone (dex), by Merck Group (4 mentions)
Dmem f12, by Thermo Fisher Scientific (4 mentions)
Putrescine, by Merck Group (3 mentions)
Triiodothyronine, by Merck Group (3 mentions)
Ascorbic acid, by Merck Group (3 mentions)
Transferrin, by Merck Group (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Ascorbate 2 phosphate, by Merck Group (2 mentions)
Selenium, by Merck Group (2 mentions)
Human recombinant ngf, by Thermo Fisher Scientific (2 mentions)
Charcoal stripped fbs, by Thermo Fisher Scientific (2 mentions)
Matrigel, by Corning (2 mentions)
Exoquick tc exosome isolation kit, by System Biosciences (2 mentions)
Lugol s iodine, by Merck Group (1 mentions)
9 cis retinoic acid, by Merck Group (1 mentions)
Indomethacin, by Merck Group (1 mentions)
Retinol, by Merck Group (1 mentions)
In cell analyzer 1000, by GE Healthcare (1 mentions)
15 protocols using thyroxine
1
Investigating Thyroid Hormone Modulators
2
Assessing OPC Survival with PDE7-GSK3 Inhibitors
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Purified OPCs were placed on coverslips coated with poly-L-lysine (Sigma) and laminin (Engelbreth-Holm-Swarm murine sarcoma; Sigma; 2 × 104 cells/well) and they were cultured with dual PDE7-GSK3 or GSK3 inhibitors (VP3.15 or TDZD8; 1 μM) in a previously described serum-free differentiation medium78 (link), consisting of BME:F12 (1:1; Invitrogen) supplemented with 100 μg/ml transferring (Sigma), 20 μg/ml putrescine (Sigma), 12.8 ng/ml progesterone (Sigma), 10.4 ng/ml sodium selenite (Sigma), 25 μg/ml insulin (Sigma), 0.8 μg/ml thyroxine (Sigma), 0.6% glucose (Normapur), 6.6 mM glutamine (Invitrogen) and 100 U/mL penicillin, 0.1 mg/ml streptomycin and 0.25 μg/ml Amphotericin B antibiotic anti-mycotic solution (Sigma). After 2 DIV, they were fixed with 4% paraformaldehyde (PFA) and subjected to immunocytochemistry for active caspase 3 (Casp3; 1:200; Abcam; Cat#ab13847) and the OPC marker A2B5 (1:10; Hybridoma Bank). 10–20 microphotographs from each coverslip were taken randomly with an In Cell Analyzer 1000 (GE-HealthCare) and the number of Casp3+-A2B5+ double positive cells was counted using the software In Cell Analyzer 1000 Workstation (GE-HealthCare). Data were expressed as a mean ratio of double positive cells for Casp3 and A2B5 with respect to the total number of A2B5+ cells ± SEM for each condition in at least three independent experiments.
3
Quantitative LC-MS/MS Thyroxine Assay
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Chromatography solvents water, acetonitrile, methanol, ammonium hydroxide, formic acid, and Centrifree YM-30 filters were supplied by Merck (Darmstadt, Germany). Thyroxine was obtained from Sigma-Aldrich (St Louis, MO; USA) (IRMM 468 European Commission Certified). The internal standard (IS) was 13C6-Thyroxine obtained from Toronto Research Chemicals T425602 (Toronto, ON, Canada).
4
Isolation and Culture of Mouse Hepatocytes
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Mouse primary hepatocytes (MPH) were freshly isolated using a two-step collagenase digestion method as previously described22 (link). MPH were cultured in William's E medium supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin, 0.1 μmol/L dexamethasone, and 0.1 μmol/L thyroxine (Sigma–Aldrich, St. Louis, MO, USA). All cells were cultured in a humidified cell culture incubator (Thermo Fisher) at 37 °C with 5% CO2.
5
Chondrogenic and Hypertrophic Differentiation of BMSCs
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For chondrogenic and hypertrophic differentiation,14 (link) 1 × 106 BMSCs were placed in polypropylene tubes, centrifuge at a speed of 500 g for 20 min. Then remove the medium and add 500 μL of low-glucose DMEM supplemented with 10−7 M dexamethasone (Sigma–Aldrich), 1%(v/v) ITS (Sigma–Aldrich), 50 μM ascorbate-2-phosphate (Sigma–Aldrich), 1 mM sodium pyruvate, 50 μg/ml proline (Sigma–Aldrich) and 20 ng/ml TGFβ3. Change the medium three times a week and culture for 14d. Then, the serum-free chondrogenic medium was replaced with hypertrophic medium supplemented with 50 nM thyroxine (Sigma–Aldrich), 10−8 M dexamethasone, 250 μM ascorbate-2-phosphate, 1 mM sodium pyruvate and 50 μg/ml proline. Change the medium three times a week and culture for another 14d.
6
Oligodendrocyte Differentiation Assay
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The preparation of mouse oligodendrocyte-lineage OL1 cells was described previously15 (link). In the differentiation assay, OL1 cells (2.0 × 104 cells) were cultured on a poly-L-ornithine (#P3655, Sigma-Aldrich)-coated 35-mm plastic dish in Knock-out DMEM/F-12 (#21331-020, Thermo Fisher Scientific) supplemented with 1 × GlutaMAX supplement (#35050061, Thermo Fisher Scientific), 1 × StemPro neural supplement (#A1050801, Thermo Fisher Scientific), 10 μg/ml PDGF-AA (#165-25541, Wako Pure Chemical), 10 nM biotin (#B4501, Sigma-Aldrich), and 30 ng/ml thyronine (#T2752, Sigma-Aldrich), and 30 ng/ml thyroxine (#T2376, Sigma-Aldrich). On the 10th day of the culture, differentiation from oligodendrocyte precursor cells to oligodendrocytes was assessed was described previously14 –16 (link). Briefly, cells were fixed and stained with anti-NG2 (a specific marker for oligodendrocyte precursor cells) and anti-MBP (a marker for matured oligodendrocytes), and the observed with a standard fluorescence microscope (Biozero BZ8000, Keyence). Differentiation from oligodendrocyte precursor cells to oligodendrocytes was expressed as the ratio of MBP-positive cells to NG2-positive cells.
7
Preparation of Chemical Stock Solutions
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Fenofibrate and gemfibrozil were obtained from Tokyo Chemical Industry (Tokyo, Japan). Methimazole, propylthiouracil, thyroxine, and fatostatin were obtained from Sigma (St. Louis, MO, USA). Stock solutions of these chemicals were prepared by dissolving in dimethyl sulfoxide (Nacalai Tesque, Kyoto, Japan). 2-Phenoxyethanol was obtained from Wako Chemical (Osaka, Japan).
8
Radioisotope-Based Thyroid Hormone Assay
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Nonradioactive 3,3’-diiodothyronine (3,3’-T2), 3,3’,5’-triiodothyronine (rT3), 3,3’,5-triiodothyronine (T3) and thyroxine (T4) were purchased from Sigma-Aldrich (Zwijndrecht, the Netherlands). [125I]-3,3’-T2, [125I]-rT3, [125I]-T3, [125I]-T4, non-radioactive and [125I]-labeled T3 sulfate (T3S) and T4 sulfate (T4S) were prepared as previously described (23 (link)). Probenecid and lesinurad (Sigma-Aldrich) were dissolved in 0.15N NaOH and DMSO, respectively.
9
Isolation and Characterization of HUVECs
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Human umbilical vein endothelial cells (HUVECs) (C2517A, Lonza, Walkersville, MD, USA) were cultured in endothelial growth medium (EGM-2) purchased from Lonza Bioscience (Walkersville, MD, USA). The HUVECs were maintained in a humidified atmosphere of 95% air and 5% CO2 at 37 °C. Norepinephrine, acetylcholine, triiodothyronine (T3), thyroxine (T4), insulin-like growth factor (IGF), cortisol, aldosterone, insulin, glucagon, 5α-dihydrotestosterone (DHT), 17β-estradiol (E2), progesterone (P4), estrone (E1), estriol (E3), fulvestrant, N(ω)-nitro-L-arginine methyl ester (L-NAME), 1,2-Bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid tetrakis(acetoxymethyl ester) (BAPTA-AM), GW4869, and dimethyl sulfoxide (DMSO) were purchased from Sigma Aldrich (St. Louis, MO, USA). An ExoQuick-Tc exosome isolation kit was purchased from System Biosciences (Palo Alto, CA, USA). The polyclonal antibody against APE1/Ref-1 was obtained from MediRedox, Inc. (Daejeon, Korea) and, the anti-CD63 antibody was obtained from Biobyt (Cambridge, UK). The monoclonal antibody against anti-heat shock protein-70 (HSP70, C92F3A-5) was obtained from Enzo Life Science (Farmingdale, NY, USA), the anti-CD9 (C-4) antibody was obtained from Santa Cruz Biotechnology (Dallas, TX, USA) and anti-ALG-2 interacting protein X (Alix, 3A9) antibody was obtained from Thermo-Fisher Scientific Inc (Waltham, MA, USA).
10
Myelination of iPSC-derived Nociceptors
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Schwann cell co-cultures were prepared as previously described (Clark et al., 2017 (link)). In brief, 30,000 rat Schwann cells were added to 30 DIV iPSC-derived nociceptors in Schwann cell basal medium [DMEM/F12 (ThermoFisher), 5 mg/ml insulin (Sigma), 100 mg/ml transferrin (Millipore), 25 ng/ml recombinant-human NGF (Peprotech), 25 ng/ml Selenium (Sigma), 25 ng/ml thyroxine (Sigma), 30 ng/ml progesterone (Sigma), 25 ng/ml triiodothyronine (Sigma) and 8 mg/ml putrescine (Sigma)]. Cells were either maintained in this medium where ‘non-myelinating’ conditions were required, or myelination was induced one week after Schwann cell addition by exposing the cells to myelination medium ((N2 medium, 1:300 phenol-free Matrigel (Corning), 5% charcoal-stripped FBS (ThermoFisher), 25 ng/ml recombinant- human NGF (Peprotech), 50mg/ml ascorbic acid (Sigma)). Myelinating co-cultures were maintained for a further 5 weeks before fixation for ICC analysis.
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