Rotenone

Manufactured by Selleck Chemicals

Sourced in United States

Rotenone is a naturally occurring compound commonly used as an insecticide, piscicide, and pesticide. It is extracted from the roots of certain tropical plants. Rotenone is effective in controlling a variety of insect pests and fish species.

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Lab products found in correlation

11 protocols using rotenone

Measuring Cellular Bioenergetics with Seahorse

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The XF^e Extracellular Flux Analyzer (Seahorse Bioscience, Billericay, Massachusetts, USA) was employed to detect the ECAR and OCR. The ECAR was detected using the XF Glycolysis Stress Test Kit (Seahorse Bioscience), while OCR was detected using XF Cell Mito Stress Test Kit. Cells were seeded in a matched microplate at a density of 1×10⁴. In the detection of the ECAR, after performing a measurement at baseline, glucose, oligomycin (an inhibitor of oxidative phosphorylation; Selleck, Wuhan, China), and 2-deoxyglucose (an inhibitor of glycolysis; Selleck) were added to each well at specific time points. In the detection of the OCR, oligomycin, FCCP (a reversible inhibitor of oxidative phosphorylation; Selleck), and rotenone (an inhibitor of mitochondrial complex I; Selleck) plus antimycin A (an inhibitor of mitochondrial complex III; Selleck) were added in the proper order. The data were collected and analyzed using the XF-96 wave software(Seahorse Bioscience). All data were normalized based on the concentration of protein.

Pan C., Liu Q, & Wu X. (2019). HIF1α/miR-520a-3p/AKT1/mTOR Feedback Promotes The Proliferation And Glycolysis Of Gastric Cancer Cells. Cancer Management and Research, 11, 10145-10156.

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Targeting Cellular Pathways with Compounds

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A939572 (Tocris 4845), GSK2194069 (Tocris 5303), PF-05175157 (Tocris 5790), and rotenone (Selleck Chemicals S2348) were used at the indicated concentrations.

Lien E.C., Westermark A.M., Zhang Y., Yuan C., Li Z., Lau A.N., Sapp K.M., Wolpin B.M, & Vander Heiden M.G. (2021). Low glycemic diets alter lipid metabolism to influence tumor growth. Nature, 599(7884), 302-307.

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Modeling Parkinson's Disease in SH-SY5Y Cells

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Human neuroblastoma SH-SY5Y cells (#CL-0208) were purchased from Procell (Wuhan, China) and cultured in Minimum Eagle Medium (MEM, Procell, Wuhan, China) supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin. Cells were incubated at 37 °C and 95% humidity with 5% CO₂. To contribute PD cell culture model in vitro, SH-SY5Y cells were seeded in 1 ml media at a density of 1 × 10⁵ per well in a 12-well plate for approximately 24 h., then were treated with indicated drugs respectively. In control group, cells were exposed to medium (containing 0.1% dimethyl sulfoxide). In model group, cells were treated with 1 µmol/L (µM) rotenone (#S2348, Selleck, Houston, USA) with or without 5 µmol/L (µM) liproxstatin-1 (#S7699, Selleck, Houston, USA). After being treated with 24 h, the cells were digested and washed with phosphate buffer saline and subjected to further treatment and analysis.

Wang M., Li T., Gao R., Zhang Y, & Han Y. (2023). Identifying the potential genes in alpha synuclein driving ferroptosis of Parkinson’s disease. Scientific Reports, 13, 16893.

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Extracellular Metabolic Profiling of Osteosarcoma Cells

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The XFe Extracellular Flux Analyzer (Agilent Technologies, Inc.) was used to detect the ECAR and OCR in the present study. The ECAR was detected using an XF Glycolysis Stress Test kit (Agilent Technologies. Inc.), and the OCR was detected using an XF Cell Mito Stress Test kit. A total of 1×10⁴ MG63 and Saos-2 cells were seeded in matched 96-well microplates. Following baseline measurements, 30 mM glucose, 2 µM oligomycin (an inhibitor of oxidative phosphorylation; Selleck Chemicals) and 10 mM 2-DG (an inhibitor of glycolysis; Selleck Chemicals) were added to each well at the specified time points to detect the ECAR. Similarly, 2 µM oligomycin, 5 µM FCCP (a reversible inhibitor of oxidative phosphorylation; Selleck Chemicals) and 1 µM rotenone (an inhibitor of mitochondrial complex I; Selleck Chemicals) plus 1 µM antimycin A (an inhibitor of mitochondrial complex III; Selleck Chemicals) were added in the appropriate order to detect the OCR and ECAR according to the operating steps described in a previous study (20 (link)). The data were collected and analyzed using XF-96 Wave software version 2.1 (Agilent Technologies. Inc.). The data were normalized based on the protein concentration.

Yan X., Yang C., Hu W., Chen T., Wang Q., Pan F., Qiu B, & Tang B. (2020). Knockdown of KRT17 decreases osteosarcoma cell proliferation and the Warburg effect via the AKT/mTOR/HIF1α pathway. Oncology Reports, 44(1), 103-114.

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Ginsenoside Rg1 Attenuates Endothelial Dysfunction

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Ginsenoside Rg1 was obtained from Nanjin Jingzhu Biotechnology Company. Assay kits for Cell Counting Kit-8, NO nitrate reductase, dihydroethidium (DHE), MDA, JC-1, and MitoSOX were obtained from Beyotime Biotechnology (Shanghai, China). Antibodies against calpain-1 and β-actin were obtained from Proteintech (Wuhan, China). Antibodies against Drp1, Mfn2, PP2A, Ser¹¹⁷⁷ eNOS phosphorylation and endothelial nitric oxide (eNOS) were obtained from ABclonal (Wuhan, China). Phenylephrine (PE), acetylcholine (Ach), polyethylene glycol-catalase (PEG-Cat), 5-hydroxydecanoate (5-HD) and okadaic acid (OKA) were obtained from Sigma Aldrich (Shanghai, China). MDL-28170, L-NAME, acetylcysteine (N-acetyl-l-cysteine, NAC), TEMPOL and rotenone were obtained from Selleck (Houston, TX, USA).

Zhao F., Lu M, & Wang H. (2022). Ginsenoside Rg1 ameliorates chronic intermittent hypoxia-induced vascular endothelial dysfunction by suppressing the formation of mitochondrial reactive oxygen species through the calpain-1 pathway. Journal of Ginseng Research, 47(1), 144-154.

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Metabolic Profiling of CD8+ T Cells

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OCRs and ECARs were measured using a Seahorse XFe96 extracellular flux analyzer (Agilent Technologies). Briefly, OCR of CD8⁺ T cells were analyzed under basal conditions and after sequential injections of 1 μM oligomycin A (APExBIO Technology), 2 μM carbonyl cyanide p-trifluoromethoxyphenylhydrazone (Selleckchem), and 0.1 μM rotenone (Selleckchem) with 1 μM antimycin A (Abcam). ECAR was analyzed under basal conditions and after sequential injections of 10 mM glucose (Sigma-Aldrich), 1 μM oligomycin A, and 50 mM 2-deoxyglucose (Sangon Biotech).

Zhai X., Liu K., Fang H., Zhang Q., Gao X., Liu F., Zhou S., Wang X., Niu Y., Hong Y., Lin S.H., Liu W.H., Xiao C., Li Q, & Xiao N. (2021). Mitochondrial C1qbp promotes differentiation of effector CD8+ T cells via metabolic-epigenetic reprogramming. Science Advances, 7(49), eabk0490.

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Ferroptosis and Cell Death Induction

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Alkaloid natural product library was obtained from TargetMol (TargetMol, Shanghai, China, L6110). RSL3 (Selleckchem, Houston, TX, S8155) and erastin (Selleckchem, S7242) were used to induce ferroptosis. Staurosporine (Selleckchem, S1421), rapamycin (Selleckchem, S1039), rotenone (Selleckchem, S2348)/H₂O₂ (Sigma-Aldrich, MO, 31642) were used to induce apoptosis, autophagy, and necrosis, respectively. Ferrostatin-1 (Selleckchem, S7243)/liproxstatin-1 (Selleckchem, S7699), Z-VAD-FMK (Selleckchem, S8102), 3-methyladenine (Selleckchem, S2767), necrostatin-1 (Selleckchem, S8037) were used to inhibit ferroptosis, apoptosis, autophagic cell death and necrosis, respectively. N-acetylcysteine (NAC) (Selleckchem, S1623) is an antioxidant that removes ROS. BBIQ compounds used in the study were all obtained from Selleckchem which include fangchinoline (S3611), cepharanthine (S4238), berbamine (S9141), dauricine (S9295), daurisoline (S9150), neferine (S5144), isoliensinine (S9247), liensinine (S9411).

Fan Y., Zhang Y., Shi K., Cheng S., Pei D, & Shu X. (2022). Identification of a group of bisbenzylisoquinoline (BBIQ) compounds as ferroptosis inhibitors. Cell Death & Disease, 13(11), 1000.

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Exploring Ferroptosis Regulators in Cells

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Regents used in this study including Rotenone (no. S2348, Selleck), TW-37 (no. 20999, Cayman Chemical), Rapamycin (no. HY-10219, MCE),Dorsomorphin (no. 11967, Cayman Chemical), 5-Fluorouracil (no. HY-90006, MCE), Shikonin (no. HY-N0822, MCE), FIN56 (no. HY-103087, MCE), Polyphyllin VI (no. S9302, Selleck), RSL3 (no. 19288, Cayman Chemical), Z-VAD-FMK (no. HY-16658B, MCE), Necrostatin-1 (no. HY-15760, MCE), Ferrostatin-1 (no. HY-100579, MCE), Erastin (no. S7247, Selleck), iFSP1 (no. 29483, Cayman Chemical), Sorafenib (no. 10009644, Cayman Chemical), BSO (no. S9728, Selleck), Acetylcysteine (no. HY-B0215, MCE), GSH (no. HY-D0187, MCE), Cisplatin (no. HY-17394, MCE), Abrine (no. HY-N1436, MCE) were obtained from the indicated vendors.
The antibodies were as follows: SPTBN2 (Abcam, no. ab238055 for IF, IHC, WB; no.ab264178 for IP), SLC7A11 (Proteintech, 26864-1-AP for WB; Abcam, no.ab307601 for IF, IHC, WB, IP, Flow Cyt), Arp1 (Abcam, no.ab203833), Flag (Abcam, no.ab205606), HA (Abcam, no.ab9110); Tubulin (Proteintech, no.11224-1-AP), Vinculin (Proteintech, no.66305-1-Ig), Na⁺/K⁺ATPase (Beyotime, no.AF1864).

Deng J., Lin X., Qin J., Li Q., Zhang Y., Zhang Q., Ji C., Shen S., Li Y., Zhang B, & Lin N. (2024). SPTBN2 suppresses ferroptosis in NSCLC cells by facilitating SLC7A11 membrane trafficking and localization. Redox Biology, 70, 103039.

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Lactate Inhibitors Modulate Protein Lactylation in Toxoplasma gondii

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Before complete release of T. gondii RH tachyzoites from Vero cells, inhibitors were added at concentrations of 0, 5, and 20 mM, to the cells at 37 °C for 4 h. After 4 h, tachyzoites of T. gondii RH were centrifuged and rinsed with cold sterile 1× PBS buffer after 2 h. Total protein was extracted and protein lactylation of T. gondii RH was examined through Western blotting. Lactate inhibitors including sodium dichloroacetate (DCA), sodium oxalate, and rotenone were purchased from Selleck (Catalog Nos. S8615, S6871, and S2348, Selleck, Houston, TX).

Yin D., Jiang N., Cheng C., Sang X., Feng Y., Chen R, & Chen Q. (2022). Protein Lactylation and Metabolic Regulation of the Zoonotic Parasite Toxoplasma gondii. Genomics, Proteomics & Bioinformatics, 21(6), 1163-1181.

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Comprehensive Cell Death Pathway Modulation

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Necrostatin-1 (S8037), ferrostatin-1 (S7243), Z-VAD-FMK (S7023), RSL3 (S8155), erastin (S7242), deferoxamine (S5742), eugenol (S4706), necrostatin-1 (S8037), necrosulfonamide (S8251), and rotenone (S2348) were bought from Selleck Chemicals. Buthionine sulfoximine (BSO, B2515), diethyl butylmalonate (DBM, 112038), NaN3 (S8032), MG-132 (M7449), CHX (C7698), and DMSO (156914) were obtained from Sigma-Aldrich. Antimycin (1397-94-0) was purchased from Santa Cruz Biotechnology. Phosphate-buffered saline (10010023) and Opti MEM medium (51985034) were obtained from Gibco BRL. Primary antibodies against BRD7 (ab56036), phospho-p53 (ab33889), p53 (ab131442), SLC25A28 (ab80467), VDAC (ab14734), Lamin B (ab16048), and β-actin (ab179467) were obtained from Abcam Technology. Anti-mouse IgG (7076) and anti-rabbit IgG (7054) were bought from Cell Signaling Technology.

Zhang Z., Guo M., Shen M., Kong D., Zhang F., Shao J., Tan S., Wang S., Chen A., Cao P, & Zheng S. (2020). The BRD7-P53-SLC25A28 axis regulates ferroptosis in hepatic stellate cells. Redox Biology, 36, 101619.

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