Rotenone, Antimycin A, goat anti-mouse IgG-HRP, and goat anti-mouse IgG-FITC were bought from Santa Cruz (Santa Cruz Biotechnology, Inc., CA, USA). Anti-Peroxiredoxin-3 antibody was purchased from Abcam (Abcam, Cambridge, MA, USA). Superoxide dismutase (SOD) was bought from Sigma (Sigma-Aldrich, MO, USA). Rat cleaved caspases 3 and 9 ELISA kits were purchased from Chenglin (Chenglin Biotechnology, Beijing, China). TUNEL assay kit was purchased from Boster (Boster, Wuhan, China). FRTL, diethyldithiocarbamate (DETC), MitoSOX Red, MTT, LDH kit, and thyroid stimulating hormone (TSH) were purchased as previously described [2 (link)].
Rotenone
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Rotenone is an organic compound that is extracted from the roots of certain plants. It is a widely used research tool in cell biology and neuroscience due to its ability to inhibit mitochondrial complex I, an essential component of the electron transport chain. Rotenone is primarily utilized in laboratory settings to study cellular processes and mechanisms related to energy metabolism and oxidative stress.
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Lab products found in correlation
Antimycin a, by Santa Cruz Biotechnology (3 mentions)
Oligomycin, by Santa Cruz Biotechnology (3 mentions)
Antimycin, by Santa Cruz Biotechnology (3 mentions)
Adenosine diphosphate adp, by Merck Group (2 mentions)
6 diazo 5 oxo l norleucine don, by Merck Group (2 mentions)
Doxycycline dox, by Merck Group (2 mentions)
Fluoro carbonyl cyanide phenylhydrazone (fccp), by Santa Cruz Biotechnology (2 mentions)
Sodium dichloroacetate dca, by Merck Group (2 mentions)
Goat anti mouse igg fitc, by Santa Cruz Biotechnology (1 mentions)
Goat anti mouse igg hrp, by Santa Cruz Biotechnology (1 mentions)
Anti peroxiredoxin 3 antibody, by Abcam (1 mentions)
Tunel assay kit, by Boster Bio (1 mentions)
Superoxide dismutase sod, by Merck Group (1 mentions)
Phgdh inactive, by Cayman Chemical (1 mentions)
Cb 839, by Focus Biomolecules (1 mentions)
Flagellin, by InvivoGen (1 mentions)
L4130, by Merck Group (1 mentions)
Adenosine triphosphate (atp), by InvivoGen (1 mentions)
Monosodium urate msu crystals, by Merck Group (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
10 protocols using rotenone
1
Oxidative Stress Signaling Pathway
2
Metabolic Modulation in Cancer Research
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The following drugs were used: 6‐Diazo‐5‐oxo‐l ‐norleucine (Don, D2141, Sigma‐Aldrich); Sodium dichloroacetate (DCA, 347795, Sigma‐Aldrich); NCT‐502 and PHGDH inactive (19716 and 19717, Cayman); Doxycycline (Dox, D9891, Sigma‐Aldrich), Oligomycin (sc‐203342, Santa Cruz Biotechnology); FCCP (sc‐203578, Santa Cruz Biotechnology); Antimycin (sc‐202467, Santa Cruz Biotechnology); Rotenone (sc‐203242, Santa Cruz Biotechnology); Cyt.C (C2037, Sigma‐Aldrich) CB‐839 (10‐4556, Focus Biomolecules); Adenosine diphosphate (ADP, A2754, Sigma‐Aldrich).
3
Inflammasome Activation in Macrophages
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As shown in Figure 1 B, macrophages (i.e., BMDMs and PMA-treated THP-1 cells) were seeded on a 12-well plate (1.0 × 106 cells per well) in RPMI 1640 containing 10% FBS and antibiotics, and treated with LPS (1 μg/mL; L4130, Sigma-Aldrich Co., Burlington, MA, USA) for 3 h [5 (link),9 (link),10 (link)]. The LPS-primed cells were replaced with RPMI 1640 media (350 μL per well) containing an inflammasome trigger with maltol (ESFOOD, #186785643, Gunpo-si, Gyeonggi-do, Korea). The inflammasome triggers are as follows: ATP (5 mM, InvivoGen, San Diego, CA, USA) for 1 h, nigericin (NG, 40 μM, Tocris Bioscience, Bristol, UK) for 1 h, monosodium urate crystals (MSU, 400 μg/mL, Sigma-Aldrich Co., Burlington, MA, USA) for 3 h, flagellin (500 ng/mL; Invivogen, San Diego, CA, USA) with Lipofectamine 2000 (10 μL/mL, Invitrogen, Carlsbad, CA, USA) for 3 h, dsDNA (1 μg/mL) with jetPRIMETM (2 μL/mL, Polyplus-transfection Inc., Illkirch, France) for 1 h, LPS (2 μg/mL, Sigma-Aldrich Co., Burlington, MA, USA) with FuGENE® HD (2.5 μL/mL, Roche, Penzberg, Germany) for 6 h, and rotenone (160 μM, Santa Cruz Biotechnology, Dallas, TX, USA) for 6 h [24 (link),25 (link)].
4
Ferroptosis Inducers and Inhibitors Protocol
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Liproxstatin-1, Z-VAD-FMK, necrostatin-1, erastin, (1S,3R)-RSL3 (RSL3), ferroptosis inducer (FIN56), 20S proteasome inhibitor, cycloheximide, deferoxamine, eugenol, and necrosulfonamide were obtained from Selleck Chemicals. Rotenone, diethyl butylmalonate, antimycin, and NaN3 were purchased from Santa Cruz Biotechnology. Anti-DACH1 antibody, anti-p53 antibody, anti-p53 (phospho S392) antibody, anti-SLC25A37 antibody, anti-Hsp90 antibody, anti-Voltage dependent anion channel protein antibody, anti-lamin B antibody, and anti-beta actin antibody were obtained from Abcam Technology. Anti-mouse IgG and anti-rabbit IgG were bought from Cell Signaling Technology.
5
Metabolic Modulation in Disease Models
6
GSSSG Synthesis and Application
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Dimethyl sulfoxide (DMSO) was purchased from Wako Pure Chemicals (Osaka, Japan). Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) was purchased from Abcam (Cambridge, UK). Antimycin A, rotenone and sulfasalazine (SASP) were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Oligomycin was purchased from Alomone Labs (Jerusalem, Israel). Glutathione trisulfide (GSSSG) was synthesized as previously described [27 (link),28 ]. GSSSG was dissolved in distilled water to prepare a 1 mM stock solution until analysis. Sodium acetate, 30 mM, was added to adjust the stock solution to a pH of 5.0. The stock solution was diluted with culture medium to the indicated final concentration.
7
Synthesis and Characterization of IM176OUT05
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IM176OUT05 (IM) is synthesized as a derivative of biguanide with improved potency and pharmacokinetic properties. Rotenone was purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). Phenformin, oligomycin A, FCCP, and 4’,6-diamidino-2-phenylindole (DAPI) were purchased from Sigma (St. Louis, MO, USA). Minoxidil was purchased from Hyundai Pharm (Seoul, Republic of Korea). Metformin was purchased from Cayman Chemical (Ann Arbor, MI, USA).
8
Drosophila Genetic Manipulation and ETC Inhibition
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Wild-type flies (white Dahomey, wDAH), RNA interference (RNAi) and GAL4 driver lines were collected and cultured as in [41 (link)]. Briefly, flies were maintained on standard media (1% Drosophila agar type II (Dutscher Scientific, #789,150), 1.5% sucrose (Sigma, #S27480), 3% glucose (Sigma, #16,325), 3.5% dried yeast (Dutscher Scientific, #789,093), 1.5% maise (TRS), 1% wheat (MP Biomedicals, #0,290,328,805), 1% soy (Santa Cruz Biotechnology, #Sc-215897A), 3% treacle (Bidvest, #90028S), 0.5% propionic acid (VWR, #8.00605.2500), 0.1% Nipagin (Sigma, #H5501)), collected using CO2 anaesthesia within 24 h of eclosion and maintained at a density of 20 flies per vial at 25 °C. Female flies 2–5 days old, unless otherwise stated, were used for all experiments. UAS-levy-RNAi (CG17280, 101,523) and UAS-ND-75-RNAi (CG2286, 100,733) were obtained from the Vienna Drosophila Resource Centre (VDRC), while daughterless-GAL4 (daGAL4) was acquired from the Bloomington Drosophila Stock Centre (BDSC). ETC inhibitors, rotenone (Santa Cruz Biotechnology, #Sc-203242) and cyanide (Sigma, #60,178), were dissolved in ethanol and water and added to the fly food at a final concentration of 600 µM and 18 mM, respectively.
9
Rotenone-Induced Parkinson-like Model in NSC34 Cells
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The mouse motoneuron-like hybrid cell line NSC34 [41 (link)] was cultured in Dulbecco’s modified Eagle’s medium (DMEM, Corning, NY, USA) supplemented with 10% (v/v) fetal bovine serum (FBS), 1% (v/v) L-glutamine, 1% (v/v) penicillin/streptomycin (Sigma-Aldrich, St. Louis, MO, USA) at 37 °C in a humidified atmosphere of 5% CO2. Rotenone (Santa Cruz Biotechnologies, Dallas, TX, USA), an inhibitor of mitochondrial complex I, used to induce Parkinson-like syndrome as an experimental model in rats, was added to the culture medium at 100 nM for 24 h.
10
Mitochondrial Respiration Assay
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Culture media and supplements were purchased from Thermo Scientific, except DMEM without glucose (cat. D5030), that was from Sigma-Aldrich. Oligomycin, CCCP, rotenone, and antimycin A were from Santa Cruz Biotechnology. All the other reagents were purchased from Sigma-Aldrich, unless stated differently.
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