Mouse feed containing 300 ppm Cd was prepared by Oriental Yeast (Tokyo, Japan) with a certified diet (MF, Oriental Yeast) and CdCl2 (Fujifilm Wako Pure Chemical., Osaka, Japan). Four-week-old female C57BL/6J mice were purchased from CLEA Japan (Tokyo, Japan) and randomly assigned to control (n = 24) or Cd-exposed groups (n = 21) (approval protocol code: 14-024). These mice were maintained in the laboratory animal facility of the School of Pharmacy, Aichi Gakuin University, at 23 ± 1 °C, with 55 ± 15% relative humidity. This laboratory animal facility is checked twice a year for contamination by pathogenic microorganisms, and no pathogenic microorganisms were detected during this study period. Mice were acclimated with MF and tap water ad libitum for 1 week. All maintenance of mice and experiments with mice were performed in accordance with the guidelines established by the Animal Care and Use Committee of the School of Pharmacy, Aichi Gakuin University.
Cdcl2
Manufactured by Fujifilm
Sourced in Japan
CdCl2 is a chemical compound used in various laboratory applications. It is a white, crystalline powder that is soluble in water and other polar solvents. CdCl2 is commonly used as a precursor for the synthesis of other cadmium-based compounds and materials.
Automatically generated - may contain errors
Lab products found in correlation
C57bl 6j mice, by CLEA Japan (1 mentions)
Sb216763, by Cayman Chemical (1 mentions)
Ag1478, by Santa Cruz Biotechnology (1 mentions)
Pifithrin α, by Fujifilm (1 mentions)
Mk 2206, by Active Motif (1 mentions)
Mek1 2, by Cell Signaling Technology (1 mentions)
Ly294002, by Cayman Chemical (1 mentions)
4 aminopyridine 4 ap, by Fujifilm (1 mentions)
Ix71 microscope, by Olympus (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Insulin transferrin selenium x, by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Hk 2 cell, by American Type Culture Collection (1 mentions)
Ferrostatin 1, by Cayman Chemical (1 mentions)
1 2 dimyristoyl sn glycero 3 phospho 1 rac glycerol, by Merck Group (1 mentions)
1 2 dimyristoyl sn glycero 3 phosphocholine, by Merck Group (1 mentions)
Opti mem medium, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (1 mentions)
Necrostatin 1, by ChemScene (1 mentions)
6 protocols using cdcl2
1
Dietary Cadmium Exposure in C57BL/6J Mice
2
Investigating Notch Signaling in Cancer Cells
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CdCl2 was obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). LY294002 and SB216763 were obtained from Cayman Chemical Company (Ann Arbor, MI, USA). AG1478 and PPP were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). DAPT and pifithrin-α were obtained from Wako. MK-2206 was obtained from Active Biochem (Maplewood, NJ, USA). Antibodies against phospho-EGFR (Tyr1068), total EGFR (D38B1) XP, phospho-Akt (Thr308) (C31E5E), total Akt (pan) (C67E7), phospho-p70 S6 kinase (Thr389) (108D2), phospho-GSK-3α/β (Ser21/9) (37F11), total GSK3-α/β (D75D3) XP, cleaved Notch1 (Val1744) (D3B8), Notch1 (D1E11) XP, Jagged1 (28H8), Jagged2 (C23D2), Snail (C15D3), E-cadherin (24E10), phospho-p53 (Ser15), and MEK1/2 were obtained from Cell Signaling Technology, Inc. (Beverly, MA, USA). p53 (DO-1), Notch1 (C-20), actin (I-19), and lamin A/C (14/LaminAC) antibodies were obtained from Santa Cruz Biotechnology. The siRNAs targeted against the human Notch1 (siRNA-1: Hs_NOTCH1_3 FlexiTube siRNA, SI00119028, siRNA-2: Hs_NOTCH1_4 FlexiTube siRNA, SI00119035), Jagged1 (Hs_JAG1_5 FlexiTube siRNA, SI02780134), Jagged2 (Hs_JAG2_2 FlexiTube siRNA, SI03095764), SNAI1 (siRNA-1: Hs_SNAI1_1 FlexiTube siRNA, SI00083398, siRNA-2: Hs_SNAI1_5 FlexiTube siRNA, SI02636424), and non-target siRNA (AllStars Negative Control siRNA) were purchased from Qiagen (Hilden, Germany).
3
Ion Channel Inhibitors Evaluation
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TTX was purchased from Sankyo Co. Ltd (Tokyo, Japan). CdCl2 and 4-aminopyridine (4-AP) were purchased from Wako Pure Chemical Industries Ltd (Tokyo, Japan). All other chemicals were purchased from Sigma Co. (St. Louis. MO, USA). E-4031 was a gift from Eisai Pharmaceutical Co. (Tokyo, Japan), and chromanol 293B was a gift from Hoechst Marion Roussel (Frankfurt, Germany).
4
Cardiomyocyte Beating Rate Analysis
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The beating rates of the iPS-CMs and FCMs were analyzed using several chemical compounds. Isoproterenol ISP (ISP) (0.4, 4, 40, and 400 nM) (Tokyo Chemical Industry Co., Ltd., Tokyo, Japan), nitrendipine (NDP) (10 μM) (Wako), LBP (0.05, 0.5, and 5 μM) (Toronto Research Chemicals, North York, Canada), glycine hydrazide (GlyH) (5 μM) (Santa Cruz Biotechnology, Inc., Heidelberg, Germany), and/or CdCl2 (10 μM) (Wako) were added to the culture medium. The iPS-CMs and FCMs treated with these chemical compounds were incubated for 20 min at 37 °C in a 5% CO2 incubator, and beating of the iPS-CMs and FCMs was captured at 16 frames per second using an ORCA-ER EMCCD camera (Hamamatsu Photonics K.K., Shizuoka, Japan) connected to an IX-71 microscope (Olympus Corporation, Tokyo, Japan). Changes in beats per minute (bpm) were then analyzed.
5
Cadmium Exposure in HK-2 Cell Culture
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HK-2 cells purchased from the American Type Culture Collection (Manassas, MA, USA) were cultured in Dulbecco’s modified Eagle’s medium/Ham’s F-12 nutrient mixture (Sigma-Aldrich, St. Louis, MO, USA) containing 10% fetal bovine serum (Gibco, Grand Island, NY, USA), 25 U/mL penicillin (DS Pharm, Osaka, Japan), 25 μg/mL streptomycin (DS Pharm), 1% insulin-transferrin-selenium X (Gibco), 10 ng/mL epidermal growth factor (Sigma-Aldrich), and 5 ng/mL hydrocortisone at 37 °C in a humidified incubator with 5% CO2. The cells were cultured in test plates at a density of 250 cells/mm2 for 48 h. After discarding the culture medium, the cells were treated with Cd (CdCl2; 98.0%; Fujifilm Wako Pure Chemical Co., Tokyo, Japan) in serum-free culture medium.
6
Lipid Oxidation and Ferroptosis Assay
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Lipofectamine RNAiMAX reagent and Opti-MEM medium were obtained from Invitrogen (Carlsbad, CA, U.S.A.). RSL3 was purchased from Selleckchem (Houston, TX, U.S.A.). The mouse monoclonal antibody against β-actin, the rabbit polyclonal antibody against ACSL6 (SAB4500971), NaAsO2, heptadecanoyl CoA (17:0-CoA), and 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (14:0/14:0 phosphatidylcholine (PC)), 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (14:0/14:0 PE), and 1,2-dimyristoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (14:0/14:0 phosphatidylglycerol), were obtained from Sigma–Aldrich (St. Louis, MO, U.S.A.). The rabbit monoclonal antibody against ACSL4 (FACL4: ab155282) and the rabbit polyclonal antibody against ACSL5 (ab104892) were acquired from Abcam (Cambridge, MA, U.S.A.). Ferrostatin-1 was obtained from Cayman Chemicals (Ann Arbor, MI, U.S.A.). Necrostatin-1 was acquired from ChemScene LLC (Monmouth Junction, NJ, U.S.A.). CdCl2 was purchased from Wako Chemicals (Osaka, Japan). The rabbit polyclonal antibody against ACSL1 (H-65), the mouse monoclonal antibody against ACSL3 (H-9), and DPQ were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, U.S.A.). Z-VAD-FMK (z-VAD) was obtained from the Peptide Institute, Inc. (Osaka, Japan).
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