Hinokitiol (469521) was purchased from Sigma (St. Louis, MO, USA). Stock solutions of 10 mM Hinokitiol was dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich) and stored at -80°C. Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies Inc. (Kumamoto, Japan). Annexin V-FITC/propidium iodide (PI) apoptosis straining kit was purchased from BD company (American). The mRFP-GFP-LC3 adenovirus construct was obtained from Hanbio Inc. (Shanghai, China). Hochest33258 was purchased from Kaiji Biology (Jiangsu, China). All the antibodies used were as follows: P62 (#88588), Beclin-1 (#3495), caspase-3 (#9664), Bcl-2 (#4223), p-mTOR (#D9C2), and p-GSK3β (Ser9) (#9336) were obtained from Cell Signaling Technology (Beverly, USA). P21 (#55643) was purchased from BD PharMingen (New York, USA). GAPDH (60004) was purchased from Proteintech Group, Inc. (USA).
Hinokitiol
Manufactured by Merck Group
Sourced in United States
Hinokitiol is a naturally occurring compound derived from the essential oil of certain plants. It functions as a chelating agent, capable of binding to and transporting metal ions. This property makes Hinokitiol a versatile compound with potential applications in various laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Horseradish peroxidase hrp conjugated donkey anti rabbit immunoglobulin g igg, by Cytiva (2 mentions)
Sheep anti mouse igg, by Cytiva (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
P gsk3β ser9, by Cell Signaling Technology (1 mentions)
Bcl2 4223, by Cell Signaling Technology (1 mentions)
P mtor d9c2, by Cell Signaling Technology (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Citral, by Merck Group (1 mentions)
Menthol, by Merck Group (1 mentions)
Geraniol, by Merck Group (1 mentions)
Linalool, by Merck Group (1 mentions)
Camphor, by Merck Group (1 mentions)
Fluconazole, by Merck Group (1 mentions)
Eugenol, by Merck Group (1 mentions)
Thymol, by Merck Group (1 mentions)
Carvacrol, by Merck Group (1 mentions)
3 3 5 5 tetramethylbenzidine (tmb), by Merck Group (1 mentions)
Holo tf, by Merck Group (1 mentions)
Anti phospho pi3k, by Cell Signaling Technology (1 mentions)
13 protocols using hinokitiol
1
Hinokitiol-Induced Autophagy and Apoptosis
2
Constitutively Active AKT in Murine Cancer
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Hinokitiol (purity: 99%), rhodamine 123 (Rho-123), 5-Fluorouracil (5-FU) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (Sigma Aldrich, St. Louis, MO, USA). Dr. Chiau-Yuang Tsai (Department of Molecular Immunology, Osaka University, Japan) provided the constitutively active AKT plasmid 18 (link). The B16F10 (mouse melanoma) 18 (link) and CT26 (mouse colon cancer) 19 (link) cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 1% antibiotics (100 units/mL penicillin and 100 μg/mL streptomycin), 2mM l-glutamine and 10% fetal bovine serum (FBS). The C57BL/6 (B16F10 mouse tumor model) and BALB/c (CT26 mouse tumor model) female mice were purchased from the National Laboratory Animal Center of Taiwan. The experimental protocol adhered to the rules of the Animal Protection Act of Taiwan and was approved by the Laboratory Animal Care and Use Committee of the National Sun Yat-sen University.
3
Antifungal Effects of Natural Compounds
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C. albicans (ATCC 90028, 90029), C. glabrata (ATCC 90030) and C. parapsilosis (ATCC 90018) were purchased from the American Type Culture Collection. C. guilliermondii (KCMF 20104), C. parapsilosis (KCMF 20154), fluconazole-resistant C. albicans (KCMF 20017), fluconazole-resistant C. glabrata (KCMF 20161) and fluconazole-resistant C. tropicalis (KCMF 20197) were obtained from the Korean Collection of Medical Fungi. For susceptibility to fluconazole, C. albicans, C. guilliermondii, C. parapsilosis, C. tropicalis with MIC ≥8 μg/ml were considered to be resistant and C. glabrata with MIC ≥64 μg/ml were considered to be resistant. To prepare suspensions of budding cells, all of the strains were propagated in yeast extract peptone dextrose (YPD) medium and incubated overnight in an orbital shaker at 30°C. Stock solutions of hinokitiol, thymol, geraniol, citral, carvacrol, menthol, eugenol, linalool, camphor and fluconazole (Sigma-Aldrich, St Louis, MO, USA) were prepared utilizing dimethyl sulfoxide (DMSO) as the solvent, and stored at − 80°C until use. The final concentration of DMSO in the test solutions was less than 0.5%. Commercially available human bone marrow-derived mesenchymal stem cells (MSCs) were purchased from Lonza (Walkersville, MD, USA).
4
Synthesis and Characterization of Fe(Hino)3
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Hinokitiol was purchased from Sigma (catalog no. 469521). Fe(Hino)3 was synthesized according to the protocol described previously (26 (link)), and the purity was verified by carbon/hydrogen/nitrogen analysis (SI Appendix, Table S5 ) and inductively coupled plasma mass spectrometry measurement of iron (SI Appendix, Table S6 ). Purified human apo-Tf (catalog no. T1147), holo-Tf (catalog no. T4132), and 3,3′,5,5′-tetramethylbenzidine (catalog no. 860336) were purchased from Sigma.
5
Signaling Pathways Modulation by Hinokitiol
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Hinokitiol (product number: 469521; purity: ≥ 98.5%) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) were purchased from Sigma Chemical Company (St. Louis, MO, USA). Recombinant PDGF-BB was purchased from Pepro-Tech (Rocky Hill, NJ, USA). Anti-mouse and anti-rabbit immunoglobulin G-conjugated horseradish peroxidase (HRP) was purchased from GE Healthcare (Sunnyvale, CA, USA) and/or Jackson-Immuno Research (West Grove, PA, USA). Anti-phospho-ERK1/2 (Thr202/Tyr204), anti-phospho-AKT (Ser473), antiphospho-PI3K, and antiphospho-JAK2 monoclonal antibodies (mAbs) were purchased from Cell Signaling (Beverly, MA, USA). Anti-p53 was obtained from GeneTex Inc (Irvine, CA). The WWP-luc (p21cip/Waf1 promoter) construct (Addgene plasmid 16451) and the PG13-luc plasmid with p53 binding sites (Addgene plasmid 16642) were kindly provided by Dr. Ming Jen Hsu. The Dual-Glo luciferase assay system was purchased from Promega (Madison, WI). The Hybond-P polyvinylidene difluoride (PVDF) membrane and enhanced chemiluminescence (ECL) Western blotting detection reagent and analysis system were obtained from GE Healthcare (Sunnyvale, CA, USA). All other chemicals used in this study were of reagent grade.
6
Cell Culture Conditions and Hinokitiol Preparation
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AS-B145 cells were cultured in Minimum Essential Medium (MEM) Alpha Medium supplemented with 2 mM L-glutamine, 10% heat-inactivated fetal bovine serum and 5 μg/ml insulin at 37°C in 5% CO2. BT-474 cells were cultured in Dulbecco's modified Eagle medium and Ham's F-12 (DMEM/F12) medium (1:1) supplemented with 2 mM L-glutamine and 10% heat-inactivated fetal bovine serum (FBS) at 37°C in 5% CO2. Hinokitiol was purchased from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO) and dissolved in absolute ethanol as a stock of 100 mM.
7
Tailored Biomaterial Scaffold Fabrication
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The raw materials were gelatin (type B from bovine skin with an average molar mass of 40,000–50,000 g/mole; Sigma-Aldrich®, St. Louis, MO, USA), hyaluronic acid (molecular weight ranging within 8–10,000 kDa; Foodchemifa Co., Ltd., Tokyo, Japan), EDC (molecular weight of 191.70 g/mole; Sigma-Aldrich®, St. Louis, MO, USA), spherical micrometer-scale modified powder of DCPA (modified DCPA; REALBONE TECHNOLOGY Co., Ltd., Kaohsiung, Taiwan), and hinokitiol (purity of 99.9%; Sigma-Aldrich®, St. Louis, MO, USA). The modified DCPA powder had a particle distribution size ranging from 1 μm to 3 μm, and a powder with 98% purity was used.
8
Pharmacological Agents Protocol
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Hinokitiol (Sigma-Aldrich, Dorset, UK), Ang II (Sigma-Aldrich, Munich, Germany), ketamine (Sigma pharmaceutical industries, Menoufia, Egypt), and xylazine (Seton®, Laboratories Calier, Barcelona, Spain) were used in this study.
9
Hinokitiol and Autophagy Modulation
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Hinokitiol was purchased from Sigma-Aldrich (St. Louis, MO, USA) and dimethyl sulfoxide (DMSO) was purchased from ECHO Chemical Co. Ltd. (Taipei, Taiwan). Hinokitiol is dissolved in DMSO in 100 μM concentration. The final DMSO concentration was less than 0.1% and the highest amount of DMSO was added to the control group. Rapamycin and Bafilomycin A1 (BafA1) were obtained from Cayman Chemical (Ann Arbor, MI, USA).
10
Hinokitiol Modulates Constitutively Active AKT
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B16F10 or 4T1 cells (1 × 106 cells per well) were seeded in 6-well culture plates incubated for 24 h at 37°C, 5% CO2. Then, the cells were either transfected with 5 μg pCDNA 3.1 control plasmid or constitutively active AKT plasmid (Provided by Dr. Chiau-Yuang Tsai of the Department of Molecular Immunology, Osaka University, Japan) using Lipofectamine 2000 18 (link), 19 and then incubated for another 24 h at 37°C, 5% CO2. Afterwards, the cells were washed and treated with hinokitiol (Sigma Aldrich, St. Louis, MO, USA) for 16 h and prepared for Western blotting.
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