0.2% Acanthopanax injection (containing total flavonoids 2 mg/mL) was purchased from Heilongjiang Ussuri River Pharmaceutical Co., Ltd. (China; Approval number Z23021162), and this injection was endotoxin free for use. 3-Methyladenine was purchased from Sigma-Aldrich (USA) and 100 nmol/μL 3-Methyladenine solution was prepared as follows: 0.009 g 3-Methyladenine (formula weight 149.15) powder was dissolved in the 600 μL double distilled water and then placed in 50°C bath water.
3 methyladenine
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Italy, China, Macao, Sao Tome and Principe
3-methyladenine is a chemical compound used as a research tool in molecular biology and biochemistry laboratories. It is a modified form of the nucleic acid adenine, with a methyl group attached at the 3rd position. This compound is commonly utilized in experiments studying DNA repair mechanisms and cellular response pathways.
Automatically generated - may contain errors
Lab products found in correlation
Rapamycin, by Merck Group (43 mentions)
Chloroquine, by Merck Group (43 mentions)
Bafilomycin a1, by Merck Group (42 mentions)
Mg132, by Merck Group (21 mentions)
Cycloheximide (chx), by Merck Group (12 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (12 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (11 mentions)
N acetyl l cysteine, by Merck Group (10 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (7 mentions)
Bafilomycin a, by Merck Group (7 mentions)
Rotenone, by Merck Group (6 mentions)
Propidium iodide, by Merck Group (6 mentions)
L glutamine, by Thermo Fisher Scientific (6 mentions)
β actin, by Merck Group (6 mentions)
Caspase 3, by Cell Signaling Technology (6 mentions)
Compound c, by Merck Group (5 mentions)
P akt, by Cell Signaling Technology (5 mentions)
N acetylcysteine, by Merck Group (5 mentions)
Pepstatin a, by Merck Group (5 mentions)
Rapamycin, by LC Laboratories (5 mentions)
212 protocols using 3 methyladenine
1
Acanthopanax Injection and 3-Methyladenine Protocol
2
Cytotoxicity Assay for Ba/F3 and SA Cells
3
Modulating Melanocyte Autophagy Pathways
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Vitiligo melanocytes were treated with the AMPK inhibitor (10uM) Compound C (Sigma Aldrich, Milan, Italy) for 6 and 24 h. Normal human melanocytes were incubated with the mTORC1 inhibitor rapamycin (1 and 5 μM) (Sigma Aldrich, Milan, Italy) for 24 h. To check the role of intracellular oxidative stress in inducing autophagy process, vitiligo melanocytes were cultured in M254 containing HMGS and treated with N-acetyl-L-cystein (5 and 10 mM) (Sigma Aldrich, Milan, Italy) for 24 h. In parallel, normal human melanocytes were grown in M254 containing HMGS and exposed to 100μM of tert-butyl hydroperoxide (Sigma Aldrich) for 24 h. To inhibit autophagy, vitiligo melanocytes were grown in M254 containing HMGS and treated with the autophagy inhibitor 3-methyl adenine (5 mM) (Sigma Aldrich) for 24-48-72-96 h whereas vitiligo fibroblasts were cultured in DMEM with 10% FBS and incubated with the inhibitor 3-methyl adenine (5 mM) (Sigma Aldrich) for 24 and 48 h.
4
Preparation of Iron Chelation Agents
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Deferoxamine (Novartis Pharma SAS) was dissolved in sterile distilled water and deferasirox (Selleckchem S1712), was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 300 and 50 mmol/L, respectively. Ironomycin was a kind gift of Raphaël Rodriguez team (11 (link)), this compound was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 10 mmol/L. Other reagents: erastin (Selleckchem S7242, 10 mmol/L in DMSO), ferrostatin-1 (Selleckchem S7243, 50 mmol/L in DMSO), Q-VD Oph (SelleckChem S7311, 10 mmol/L in DMSO, 30′ pretreatment), iron(III) chloride hexahydrate (31232 Sigma Aldrich, 0.1 mol/L in water, 4 hours after treatment), reduced gluthatione GSH (Sigma Aldrich G4251, 0.1 M in PBS) H2O2 (Sigma Aldrich 216763), mafosfamide (surrogate of cyclophosphamide, Santa Cruz ChemCruz SC-211761, 10 mmol/L in saline water), gemcitabine (Sellekchem S1149, 50 mmol/L in saline water), doxorubicin (Sellekchem S1208, 20 mmol/L in DMSO), CldU (Abcam ab213715, 20 mmol/L), IdU (Abcam ab142581, 2 mmol/L), 3-methyl adenine (Merck, #189490), β-mercaptoethanol (Sigma, M3148), staurosporine (Selleckchem, S1421), ketokonazole (Sigma, UC280), Avasimibe (from Sigma, PZ0190), ibrutinib (from Sellekchem S2680), entospletinib (from Selleckchem, S7523), venetoclax (from Sellekchem, S8048), AZD-5991 (from Selleckchem, S8643), and A1155463 (from TargetMol, T6748).
5
Regulation of Intestinal Barrier Integrity
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Caco-2 cells and IEC-6 cells were obtained from the Cell Bank of the Chinese Academy of Sciences (analog no. TCHu146, Shanghai, China) and Shanghai Fuxiang Biotechnology (catalog no. CRL-1592, Shanghai, China) respectively. Cells were cultured in DMEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. Cells were maintained in a 5% CO2/95% air atmosphere at 37 °C. Recombinant TNF-α was obtained from R&D Systems (catalog no. 510-RT-010, Minneapolis, MN, USA). Bafilomycin A1 (catalog no. 189490) and 3-methyladenine (catalog no. 19-148) were purchased from EMD Millipore (Temecula, CA, USA). PP242 was purchased from Santa Cruz Biotechnology (catalog no. sc-301606, Santa Cruz, CA, USA). Claudin-2 antibody (catalog no. 51-6100, Invitrogen, Carlsbad, CA, USA), LC3B antibody (catalog no. L7543, Sigma, St. Louis, MO, USA), P62 antibody (catalog no. sc-25575, Sancta Cruz Biotechnology, Santa Cruz, CA, USA), and β-actin antibody (catalog no. 4970, Cell Signaling Technology, Danvers, MA, USA) were purchased from relevant manufacturers. Ad-mCherry-GFP-LC3B was purchased from Beyotime Biotechnology (catalog no. C3011, Shanghai, China).
6
Modulators of T cell viability
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T cells were cultured with plate bound α-CD3 (5 µg/ml) and soluble α-CD28 (2 µg/ml) with IL-2 (20 ng/ml) at 37°C in IMDM (Invitrogen) media supplemented with 10% FCS (Invitrogen) unless stated otherwise. All reagents were purchased from Sigma-Aldrich except otherwise stated. The following small molecule inhibitors were tested in threefold dilution series: β-mercaptoethanol (b-ME, 50 µM; Invitrogen), α-tocopherol (aToc, 100 µM), ascorbic acid (L-Asc, 10 µM), PD146176 (1 µM), z-VAD-FMK (100 µM; R&D Systems), Necrostatin-1 (Nec-1, 100 µM; Merck), Necrostatin-1 inactive control (Nec-1i, 100 µM; Merck), 3-methyladenine (3MA, 10 µM), U0126 (100 µM), deferoxamine (DFO, 400 µM), and Ferrostatin-1 (Fer-1, 10 µM; ChemBridge). Ebselen was purchased from Sigma-Aldrich. Cremophor EL (Sigma-Aldrich) was used to solubilize α-tocopherol in aqueous solution according to the manufacturer’s protocol. Whole splenocytes and peripheral LNs were isolated and treated in vitro with plate-bound α-CD3 (5 µg/ml; homemade), soluble α-CD28 (2 µg/ml; BioLegend), and IL-2 (20 ng/ml; homemade) at 37°C with 5% CO2.
For T cell viability analysis in vivo, mice were injected with α-CD3 (1 µg; eBioscience) i.v., and splenocytes were isolated for analysis.
For T cell viability analysis in vivo, mice were injected with α-CD3 (1 µg; eBioscience) i.v., and splenocytes were isolated for analysis.
7
Oral Squamous Cancer Cell Culture
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All chemicals were purchased from Sigma-Aldrich (St. Louis, USA) unless otherwise specified. Selective inhibitors of β2-AR (ICI118,551) and MEK1/2 (U0126) were obtained from Tocris Bioscience (Bristol, United Kingdom). 3-Methyladenine were purchased from Merck (Darmstadt, Germany).
UMSCC103 (OSCC cell line) used in this study was established at the University of Michigan under a protocol approved by the Institutional Review Board Office under the university’s regulations and described here15. The human head and neck cancer cell lines Cal-33 (oral squamous cancer cell lines) were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Cells were cultured in DMEM and RPMI (Gibco, NY, USA) supplemented with 2 mM glutamine, 100 IU/mL penicillin, 100 μg/mL streptomycin (Invitrogen, Carlsbad, CA), and 10% heat-inactivated fetal bovine serum (FBS) (Gibco, NY, USA) at 37 °C in a humidified atmosphere under 5% CO2. The cell line was kept mycoplasma-free; checking was performed every three months.
UMSCC103 (OSCC cell line) used in this study was established at the University of Michigan under a protocol approved by the Institutional Review Board Office under the university’s regulations and described here15. The human head and neck cancer cell lines Cal-33 (oral squamous cancer cell lines) were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Cells were cultured in DMEM and RPMI (Gibco, NY, USA) supplemented with 2 mM glutamine, 100 IU/mL penicillin, 100 μg/mL streptomycin (Invitrogen, Carlsbad, CA), and 10% heat-inactivated fetal bovine serum (FBS) (Gibco, NY, USA) at 37 °C in a humidified atmosphere under 5% CO2. The cell line was kept mycoplasma-free; checking was performed every three months.
8
Osteosarcoma Cell Lines: Culture and Drug Treatments
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Human osteosarcoma cell lines MG-63, Saos-2 and U-2 OS were purchased from the Cell Bank of Chinese Academy of Medical Science (Shanghai, China) and cultured in McCoy’s 5a medium (Gibco, Los Angeles, CA) containing 10% fetal bovine serum (Gibco) and ampicillin and streptomycin at 37 °C in a humidified atmosphere of 95% air and 5% CO2. All cell lines were used within 20 passages. The antibody against HSP90AA1was obtained from Proteintech. The antibodies against LC3, p62, cleaved PARP, Akt, p-Akt, mTOR, p-mTOR, JNK, p-JNK, p38, p-p38 and actin were obtained from Cell Signaling Technology. Cisplatin, doxorubicin, methotrexate, bafilomycin A1, rapamycin, LY294002 and 3-methyladenine were purchased from Sigma Aldrich (St. Louis, MO, USA).
9
Autophagy Regulation with WS2 and WSe2 Nanosheets
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WS2 and WSe2 nanosheets were obtained from XFNANO (Nanjing, China). Rapamycin and 3-methyladenine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies against LC3B and p62 were purchased from Cell Signaling Technology (Boston, MA, USA). The primary antibody β-actin was purchased from Abgent (San Diego, CA, USA). Polyvinylidene difluoride (PVDF) membranes and Western blot luminescence reagents were purchased from Millipore (Billerica, MA, USA).
10
Peptide-mediated Cell Signaling Modulation
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The TAT and TAT-vFLIP peptides were custom-synthesized with purity of more than 95% (Biomatik). Rapamycin and MG132 were purchased from Cell Signaling Technology. PU-H71 was purchased from Selleckchem. EBSS was purchased from Thermo Fisher Scientific. Recombinant TRAIL was purchased from BioLegend. Staurosporin, CCCP, 3-methyladenine, rotenone, bafilomycin A1, chloroquine, and cycloheximide were purchased from Sigma.
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