A murine proximal tubule-like epithelial cell line termed C1.1 was a kind gift of Drs. Rudolph Wuthrich and Patricia Wahl (University of Zurich, Zurich, Switzerland).14 (link) C1.1 cells were cultured in K1 medium [Dulbecco’s modified Eagle’s medium:Ham’s F12 medium (1:1 v/v) (Invitrogen, Carlsbad, CA), 1 mol/L HEPES (Invitrogen), Non-Essential Amino Acid (NEAA) medium (Invitrogen), 5 mg/mL insulin (Sigma, St. Louis, MO), 1.25 ng/mL prostaglandin E1 (Cayman Chemicals, Ann Arbor, MI), 0.0338 ng/mL 3,3,5- triiodothyronine (Sigma), 5 mg/mL hydrocortisone (Sigma), 1.73 ng/mL transferrin (Sigma), 18 ng/mL sodium selenite (Sigma), and 5% fetal bovine serum (Sigma-Aldrich, St. Louis, MO)] at 37°C, in 5% CO2. For treatment of cells, serum-supplemented media were replaced with serum-free media.
3 3 5 triiodothyronine
Manufactured by Merck Group
Sourced in United States
3,3,5-triiodothyronine is a laboratory reagent used for research purposes. It is a thyroid hormone that plays a role in various physiological processes. This product is intended for use in research applications only, and its specific functions and intended uses should be determined by the researcher based on their experimental requirements.
Automatically generated - may contain errors
Lab products found in correlation
Insulin, by Merck Group (4 mentions)
Hydrocortisone, by Merck Group (3 mentions)
Sodium selenite, by Merck Group (3 mentions)
Transferrin, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Hepes, by Thermo Fisher Scientific (2 mentions)
Prostaglandin e1, by Cayman Chemical (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
3 isobutyl 1 methylxanthine, by Merck Group (1 mentions)
Oil red o solution, by Merck Group (1 mentions)
Anti gapdh antibody, by Santa Cruz Biotechnology (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Indomethacin, by Merck Group (1 mentions)
Troglitazone, by Merck Group (1 mentions)
Dmem ham s f12, by Thermo Fisher Scientific (1 mentions)
Mem neaa, by Thermo Fisher Scientific (1 mentions)
Ab1012, by Abcam (1 mentions)
Ab5131, by Abcam (1 mentions)
Antimycin a, by Merck Group (1 mentions)
Bapta am, by Merck Group (1 mentions)
6 protocols using 3 3 5 triiodothyronine
1
Culturing Murine Proximal Tubule Cells
2
Adipogenic Differentiation Modulation
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Oil-Red O solution, 3-isobutyl-1-methylxanthine (IBMX), insulin, dexamethasone (Dex), indomethacin, 3,3′,5-Triiodothyronine (T3), and troglitazone (Trog) were bought from Sigma Chemicals Co. (St. Louis, MO, USA). Epigallocatechin-3-gallate (EGCG) was purchased from Chengdu Biopurify Phytochemicals Ltd. (Chengdu, China). Bovine serum (BS), fetal bovine serum (FBS), Dulbecco’s modified Eagle’s medium (DMEM), and penicillin-streptomycin were purchased from Gibco BRL (Grand Island, NY, USA). Anti-GAPDH antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies for pAMPK, pLKB1, PPARγ, and C/EBPα were purchased from Cell Signaling Technology (Beverly, MA, USA).
3
Mouse Tubular Epithelial Cell Culture
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Mouse tubular epithelial C1.1 cells [15] (a kind gift from the laboratory of Dr Rudolf Wuthrich, Switzerland) were cultured in DMEM:HAM's F12 (1:1 v/v) medium (Invitrogen) containing 1 M HEPES (Invitrogen), MEM NEAA (Invitrogen), 5 mg/ml insulin (Sigma), 1.25 ng/ml prostaglandin E1 (Cayman Chemicals, Ann Arbor, MI), 0.0338 ng/ml 3,3,5-triiodothyronine (Sigma), 5 mg/ml hydrocortisone (Sigma), 1.73 ng/ml transferrin (Sigma), 18 ng/ml sodium selenite (Sigma), and 5% fetal bovine serum (FBS) (Sigma-Aldrich) at 37 °C, in 5% CO 2 . For treatment of cells, serum-supplemented media were replaced with serum-free media.
4
Antibody Procurement for Histone Modifications
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3, 3',5-Triiodothyronine (>98% purity) was obtained from Sigma-Aldrich (St. Louis, MO, USA).
Antibodies against RNAPII (Cat. No. MMS-126R) and normal rabbit immunoglobulin G (15006) were purchased from Covance (Berkeley, CA, USA) and Sigma-Aldrich, respectively. Antibodies against trimethylated histone H3 lysine 4 (trimethylated H3K4; ab1012) and phosphorylated RNAPII serine 5 (phosphorylated RNAPIIS5; ab5131) were obtained from Abcam (Tokyo, Japan). Antibody against acetylated histone H4 that recognizes acetylated lysines 5, 8, 12 and 16 (06-598) was purchased from Merck Millipore (Darmstadt, Germany).
Antibodies against RNAPII (Cat. No. MMS-126R) and normal rabbit immunoglobulin G (15006) were purchased from Covance (Berkeley, CA, USA) and Sigma-Aldrich, respectively. Antibodies against trimethylated histone H3 lysine 4 (trimethylated H3K4; ab1012) and phosphorylated RNAPII serine 5 (phosphorylated RNAPIIS5; ab5131) were obtained from Abcam (Tokyo, Japan). Antibody against acetylated histone H4 that recognizes acetylated lysines 5, 8, 12 and 16 (06-598) was purchased from Merck Millipore (Darmstadt, Germany).
5
Compound Acquisition for Cell Studies
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3,5,3′-triiodothyronine (catalog no. T2877), BAPTA-AM (catalog no. A1076), antimycin A (catalog no. A8674), edelfosine (catalog no. sml0332), and chloroquine phosphate (catalog no. PHR1258) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
6
Cell Line Characterization for Oxidative Stress Research
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The HEK293, LNCaP and PNT1A cells line used in this study were purchased from Life Technologies (Carlsbad, CA, USA, Cat.# R75007 and R71407), ATCC (Manassas, VA, USA, Cat.# CRL-1740) and SIGMA (Saint-Louis, MO, USA, Cat.# 95012614), respectively. The HEK293 cell line expressing Luc UGA/Gpx4 in a stable way was generated and validated in References [15 (link),16 (link),22 (link)]. Fetal calf serum (FCS), cell culture media and supplements were purchased from Life Technologies. Transferrin, insulin, 3,5,3′-triiodothyronine, hydrocortisone, sodium selenite and sodium selenate were purchased from Sigma-Aldrich (Saint-Louis, MO, USA). Antibodies were purchased from Abcam (Cambridge, UK) (glutathione peroxidase-1 (Gpx1), #ab108429; glutathione peroxidase-4 Gpx4, #ab125066; thioredoxin reductase-1 (TxnRD1), #ab124954) and Sigma-Aldrich (thioredoxin reductase-2 (Txnrd2), #HPA003323; Actin, #A1978). NuPAGE 4-12% bis–Tris polyacrylamide gels and MOPS SDS running buffer were purchased from Life Technologies. Antiprotease inhibitor cocktail was purchased from Thermo Fisher. Selol was synthesized at the Department of Bioanalysis and Drug Analysis at the Medical University of Warsaw, as described in Reference [23 ]. A micellar solution of Selol was used (based on lecithin, water and Selol) with a declared selenium concentration of 5% (w/v).
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