The pan Bcl-2 inhibitor (−)-gossypol (>98% purity) was acquired from Tocris (Bristol, United Kingdom). Autophagy inhibitors Bafilomycin A1 (BafA1), and 3-Methyladenin (3-MA) were obtained from Sigma-Aldrich (Taufkirchen, Germany). The pan-caspase inhibitor z-Val-Ala-DL-Asp(OMe)-fluoromethylketone (z-VAD) was purchased from Bachem (Weil am Rhein, Germany). The Mcl-1 sparing Bcl-2 inhibitor ABT-737 was from Santa Cruz Biotechnology (Heidelberg, Germany) and the inductor of apoptotic cell death staurosporine (STS) was from Alexis Biochemicals (by ENZO Life Sciences, Lörrach, Germany). Chemotherapeutic gemcitabine was from Fresenius-Kabi (Bad Homburg, Germany) and chemotherapeutic cisplatin from Teva (Ulm, Germany). Lysotracker Red DND-99 was obtained from Invitrogen (Karlsruhe, Germany). All other chemicals were used in analytic grade purity from Sigma-Aldrich.
3 methyladenin 3 ma
Manufactured by Merck Group
Sourced in Germany
3-methyladenin (3-MA) is a chemical compound used in laboratory settings. It is a purine derivative with the chemical formula C6H7N5. 3-MA is commonly used as a research tool in various fields of study.
Automatically generated - may contain errors
Lab products found in correlation
Ferritin antibody for immunofluorescence, by Santa Cruz Biotechnology (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions)
Rpmi 1640, by Merck Group (2 mentions)
Ferrostatin 1, by Merck Group (2 mentions)
Secondary antibodies for western blotting, by EarthOx (2 mentions)
Staurosporine, by Alexis Biochemicals (1 mentions)
Gemcitabine, by Fresenius (1 mentions)
Cisplatin, by Teva Pharmaceuticals (1 mentions)
Bafilomycin a1 bafa1, by Merck Group (1 mentions)
Lysotracker red dnd 99, by Thermo Fisher Scientific (1 mentions)
Abt 737, by Santa Cruz Biotechnology (1 mentions)
Gossypol, by Bio-Techne (1 mentions)
Z val ala dl asp ome fluoromethylketone z vad, by Bachem (1 mentions)
Vimentin, by Proteintech (1 mentions)
Ncoa4, by Proteintech (1 mentions)
Chloroquine cq, by Merck Group (1 mentions)
Lc3 antibody, by Proteintech (1 mentions)
Di 2 pyridylketone, by Merck Group (1 mentions)
Bcl2 associated x (bax), by Wuhan Boster Biological Technology (1 mentions)
Bcl 2, by Wuhan Boster Biological Technology (1 mentions)
4 protocols using 3 methyladenin 3 ma
1
Apoptosis and Autophagy Inhibitor Assay
2
Ferroptosis Mechanism of HepG2 Cells
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HepG2 cell line was obtained from HonorGene (Changsha, China). Ferrostatin-1, MTT, 3-methyladenin (3-MA), chloroquine (CQ), RPMI-1640, and other chemicals were purchased from Sigma-Aldrich. 2-Pyridylhydrazone dithiocarbamate s-acetic acid (PdtaA) was prepared in our laboratory [27 (link)]. GPx4, xCT (SLC7A11), NDRG1, vimentin, and NCOA4 antibody were obtained from the Proteintech Group (Wuhan, China). Antibodies of E-cadherin and secondary antibodies (fluorescence-labeled for immunofluorescence) were purchased from Cell Signaling Technology (USA). Ferritin antibody for immunofluorescence was obtained from Santa Cruz Biotechnology (USA, Santa Cruz). NCOA4 antibody for immunofluorescence was purchased from Atlas Antibody (Sweden). Secondary antibodies for Western blotting were obtained from EarthOx, LLC (San Francisco, USA).
3
Ferroptosis Pathway Regulation in HepG2 Cells
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The HepG2 cell line was obtained from HonorGene; Changsha Aibiwei Biotechnology Co., Ltd. Ferrostatin-1, MTT, di-2-pyridylketone, 3-methyladenin (3-MA), RPMI-1640, and other chemicals were purchased from Sigma-Aldrich; Merck KGaA. GPx4, xCT (SLC7A11), vimentin, nuclear receptor coactivator-4 (NCOA4), and LC3 antibodies were obtained from ProteinTech Group, Inc. Antibodies against p53, caspase-8, GAPDH, Bax, and Bcl-2 were purchased from Wuhan Boster Biological Technology, Ltd. Antibodies targeting E-cadherin and secondary antibodies (fluorescence-labeled for immunofluorescence) were purchased from Cell Signaling Biotechnology, Inc. Ferritin antibody for immunofluorescence was obtained from Santa Cruz Biotechnology, Inc. NCOA4 antibody for immunofluorescence was purchased from Atlas Antibodies. Secondary antibodies for western blotting were obtained from EarthOx Life Sciences.
4
Ferroptosis Modulation in Hippocampal Neurons
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The ferroptosis inducer erastin (Cat# HY-15763, Hvcultec, Beutelsbach, Germany), Fer-1 (ferroptosis inhibitor, Cat# ab146169, Abcam, Berlin, Germany), and 3-methyladenin (3-MA [autophagy inhibitor], Sigma, Taufkirchen, Germany) were dissolved in dimethyl sulfoxide (DMSO; Sigma). Deferoxamine (DFO, ferroptosis inhibitor, Sigma) was dissolved in deionized water. Cells were seeded in 96-well plates for viability assay, 24-well plates for immunofluorescence staining, and western blot assay. To assess the optimal concentration of erastin, hippocampal HT22 neurons were exposed to the drug at 0.3, 0.4, 0.5, 0.6, 0.7, and 0.8 µM for 12 hours.
Different concentrations of Fer-1 (10 and 20 µM) and DFO (100 and 200 µM) were applied to hippocampal HT22 neurons after 12 hours of erastin treatment (0 and 5 µM). The optimal experimental conditions occurred after incubation with 20 µM for Fer-1 and 100 µM for DFO. After 6, 12, and 24 hours, the subsequent analyses were conducted.
Different concentrations of Fer-1 (10 and 20 µM) and DFO (100 and 200 µM) were applied to hippocampal HT22 neurons after 12 hours of erastin treatment (0 and 5 µM). The optimal experimental conditions occurred after incubation with 20 µM for Fer-1 and 100 µM for DFO. After 6, 12, and 24 hours, the subsequent analyses were conducted.
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