Oligomycin, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone, antimycin, and CHX were purchased from Sigma-Aldrich (St. Louis, MO). Decitabine, monensin, MG132, KX2-391, and IACS-010759 were from Selleck (Houston, TX). Recombinant human CYTL1 was purchased from OriGene Technologies (Rockville, MD). Anti-CYTL1 antibody was purchased from Abcam (Cambridge, UK). Anti-NDUFV1, anti-BRCA1, anti-Flag, anti-GAPDH, and anti-Src antibodies were from Proteintech Group (Chicago, IL). Anti-HA, anti-CD31, anti-MDM2, anti-LDHA, anti-p-LDHA (Y10), and anti-COX4 antibodies were from Cell Signaling Technology (Beverly, MA). Anti-Tubulin, anti-β-Actin, anti-myc, and p-Src (Santa Cruz) antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA). Lipofectamine 3000 and Lipofectamine RNAi MAX were purchased from Thermo Fisher Scientific (Waltham, MA). MitoCheck Complex I activity assay kit (No.700930) was purchased from Cayman Chemical (Ann Arbor, Michigan). Human CYTL1 ELISA Kit was purchased from CUSABIO Life Sciences (College Park, MD).
Monensin
Manufactured by Selleck Chemicals
Sourced in United States
Monensin is a polyether ionophore compound commonly used as a laboratory reagent. It functions as an ionophore, facilitating the transport of monovalent cations across cell membranes.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Decitabine, by Selleck Chemicals (1 mentions)
Antimycin, by Merck Group (1 mentions)
Oligomycin, by Merck Group (1 mentions)
Anti flag, by Proteintech (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Anti ha, by Cell Signaling Technology (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Mitocheck complex 1 activity assay kit, by Cayman Chemical (1 mentions)
P src, by Santa Cruz Biotechnology (1 mentions)
Kx2 391, by Selleck Chemicals (1 mentions)
Anti myc, by Santa Cruz Biotechnology (1 mentions)
Anti mdm2, by Cell Signaling Technology (1 mentions)
Anti cytl1 antibody, by Abcam (1 mentions)
Anti cox 4, by Cell Signaling Technology (1 mentions)
Anti src, by Proteintech (1 mentions)
Anti ldha, by Cell Signaling Technology (1 mentions)
Anti brca1, by Proteintech (1 mentions)
Anti tubulin, by Santa Cruz Biotechnology (1 mentions)
6 protocols using monensin
1
Mitochondrial Function and Cytokine Profiling
2
Modulation of Notch Signaling in Cancer Therapy
3
Murine Breast Cancer Cell Protocols
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Murine breast cancer cell line 4T1 and the murine macrophage cell line Raw264.7 were purchased from the Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China). Immortalized bone marrow‐derived macrophages (iBMDMs) were obtained from Shao Feng's Laboratory (National Institute of Biological Sciences, Beijing, China). 4T1 cells were cultured in RPMI‐1640, and iBMDM and Raw264.7 cells were cultured in Dulbecco's modified eagle medium (DMEM). Media was supplemented with 10% fetal bovine serum (FBS), 2 mmol/L L‐glutamine, 100 IU/mL penicillin, and 100 IU/mL streptomycin. All cell cultures were maintained at 37°C in 5% CO2. 4T1 cells were treated with the indicated concentrations of PX‐478, an inhibitor of hypoxia inducible factor‐1alpha (HIF‐1α), for 48 h, and then the cells were lysed for the detection of chemokine CX3C chemokine ligand 1 (CX3CL1) and HIF‐1α protein levels. To inhibit IL‐15Rc release, the cells were cultured alone or cocultured in the presence of the vehicle control or 4 μg/mL GM6001, a broad‐spectrum matrix metalloproteinase (MMP) inhibitor. To inhibit CX3CL1 release for further evaluation of the effect of HIF‐1α on CX3CL1 release, the cells were cultured in the presence of the vehicle control or 5 μmol/L monensin (Selleck Chemicals, LLC, Houston, TX, USA), a protein transport inhibitor.
4
Quantification of VEGFA secretion
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The cells were cultured in a 6‐well plate at a concentration of 1 × 106 cells per well for 24 h, after which the supernatant was harvested to measure VEGFA concentration using a commercially available fluorescence‐based assay kit (ARIGO, ARG81305, Taiwan, China). In addition, the cells were collected for VEGFA detection by western blot. To explore the function of Golgi in FOXM1D‐mediated exosome release, the cells were treated with 10 μm monensin (Selleck, S2324, Houston, TX, USA) for 3 h in the indicated experiment (Fig. 7D ).
5
Authentication and Culture of Human Cervical Cancer Cell Lines
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The human cervical cancer cell lines HeLa, CasKi and SiHa (Guangzhou Jennio Biotech Co. Ltd) were subjected to authentication through short tandem repeat profiling conducted by Precision Genomics Biotechnology. These cell lines were cultured in RMPI1640 medium supplemented with 10% fetal bovine serum (FBS) at 37°C, 5% CO2. Antibodies specific to phospho-β-catenin, phospho-Akt and their corresponding total were from Cell Signaling Technology. β-actin antibody was from Santa Cruz Biotechnology. Monensin, N-acetylcysteine (NAC) and lithium chloride (LiCl) were purchased from Selleckchem and were reconstituted as per manufacturer’s instructions.
6
3D Breast Cancer Cell Culture and Immunoassay
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The human breast cancer cell lines MCF-7 and T-47D were purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China), and maintained in Dulbecco's modified Eagle's medium (DMEM) with high glucose, 10% fetal bovine serum (FBS) and antibiotics (Life Technologies, Carlsbad, CA, USA), according to the guidelines. Lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Growth factor-reduced 3D Cultrex basement membrane extract (BME) was purchased from Trevigen Inc. (Gaithersburg, MD, USA). Monensin was purchased from Selleckchem.cn. Antibodies against surfactant protein C (SP-C) and aquaporin 5 (AQP5), and all secondary antibodies, were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Cell Signaling Technology Inc. (Beverly, MA, USA). The TGF-β receptor kinase inhibitor LY2109761 was purchased from Selleckchem.cn.
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