Golgicide a

Manufactured by Merck Group

Sourced in United States, Sweden

Golgicide A is a chemical compound developed for use in research laboratories. It acts as a selective inhibitor of the Golgi apparatus, a cellular organelle involved in the processing and transport of proteins within the cell. Golgicide A is employed by researchers to study the function and dynamics of the Golgi apparatus in various biological systems.

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Golgicide (2 citations)

10 protocols using golgicide a

Chitosan Synthesis and Characterization

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Cs (degree of deacetylation, 90%; viscosity, 10 mPa⋅s; average MW, ~50 kDa; utilizing shrimp and crab shells as raw materials) was purchased from Zhejiang Golden Shell Pharmaceutical Co., Ltd. (Zhejiang, People’s Republic of China). Sodium TPP (chemical reagent, 56%–60%) and acetic acid (analysis reagent, 99.5%) were supplied by Sinopharm Chemical Reagent Co. Ltd. (Shanghai, People’s Republic of China). Fluorescein isothiocyanate (FITC), rhodamine B isothiocyanate (RBITC), O-phospho-l-serine, primaquine (PRQ), golgicide A (GOL-A), sucrose, and paraformaldehyde were provided by Sigma-Aldrich Co. (St Louis, MO, USA). Vacuolin-1 (VAL-1) and wortmannin (WOT) were supplied by Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Lyso Tracker Red DND-99 was purchased from Thermo Fisher Scientific (Waltham, MA, USA). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) kit was purchased from Nanjing Jiancheng Bioengineering Institute supplied by Beyotime Biotechnology (Shanghai, People’s Republic of China). High-glucose Dulbecco’s Modified Eagle’s Medium (DMEM) was purchased from Jiangsu KeyGen Biotech Corp., Ltd (Jiangsu, People’s Republic of China). Fetal bovine serum (FBS) was purchased from Thermo Fisher Scientific.

Jiang L.Q., Wang T.Y., Webster T.J., Duan H.J., Qiu J.Y., Zhao Z.M., Yin X.X, & Zheng C.L. (2017). Intracellular disposition of chitosan nanoparticles in macrophages: intracellular uptake, exocytosis, and intercellular transport. International Journal of Nanomedicine, 12, 6383-6398.

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Pharmacological Inhibitor Preparation and Application

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Pharmacological inhibitors were prepared in recommended solvents according to the manufacturer’s instructions and stored at −20°C. Inhibitor treatments were performed with the indicated concentrations either during seeding and imaging or only during imaging (as stated in the legends). Inhibitors remained in the medium from their addition for the duration of the experiment. When performing stimulation experiments, TGF-β receptor inhibitor LY2109761 and solvent control were added to CM or control medium before stimulation. Inhibitors were purchased from the following manufacturers: eeyarestatin, tunicamycin (Cayman Chemical, #10012609 and #11445), FLI-06, golgicide A, AG1478, brefeldin A (Sigma-Aldrich, #SML0975, #G0923, #T4182, and #B7651), and LY2109761 (Cayman Chemical, #15409).

Finger A.M., Jäschke S., del Olmo M., Hurwitz R., Granada A.E., Herzel H, & Kramer A. (2021). Intercellular coupling between peripheral circadian oscillators by TGF-β signaling. Science Advances, 7(30), eabg5174.

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Modulation of Golgi Ribbon Dynamics

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Brefeldin A (BFA) (5 μM) (Sigma #B5936) and Golgicide A (10 μM) (Sigma #G0923) were added to cells in HGM for the times indicated. To unlink the Golgi ribbon cells were incubated in HGM supplemented with nocodazole (1 μg/mL) (Sigma #M1404) for 24 hours. For secretagogue stimulation cells were treated with either histamine (100 μmol/L-100μM) (Enzo #ALX-550-132-6005) or phorbol 12-myristate 13-acetate (PMA) (100 ng/mL) (Sigma #M1404) in Serum Free medium (M199 supplemented with 10 mmol/L HEPES-NaOH, pH 7.4 and 0.1 mg/mL BSA) for 30 mins unless stated. Aminoimidazole-4-carboxamide-1-b-d-ribofuranoside (AICAR) (Enzo #BML-E1330-0050) in HGM was added to cells for 24 hours in the concentration stated. 2 deoxy-D-glucose (2DG) (5.5mM) (Sigma #D6134) was added to cells in HGM for 24 hours. D-glucose (Gibco #A2494001), in the amounts indicated, was added to glucose-free DMEM (Gibco #11966025) for 24 hours.

Lopes-da-Silva M., McCormack J.J., Burden J.J., Harrison-Lavoie K.J., Ferraro F, & Cutler D.F. (2019). A GBF1-Dependent Mechanism for Environmentally Responsive Regulation of ER-Golgi Transport. Developmental Cell, 49(5), 786-801.e6.

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Molecular Signaling Pathway Inhibitors

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BML-265 was purchased from Enzo Life Sciences. Tyrphostin AG1478 and Erlotinib were ordered from Cayman chemical. DMSO, brefeldin A, Golgicide A and D-biotin were purchased Sigma-Aldrich.

Boncompain G., Gareil N., Tessier S., Lescure A., Jones T.R., Kepp O., Kroemer G., Del Nery E, & Perez F. (2019). BML-265 and Tyrphostin AG1478 Disperse the Golgi Apparatus and Abolish Protein Transport in Human Cells. Frontiers in Cell and Developmental Biology, 7, 232.

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Multifaceted Cell Death Pathway Modulation

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Indicated chemicals were obtained from companies (in parentheses): brefeldin A (Sigma-Aldrich), golgicide A (Sigma-Aldrich), AG1478/tyrphostin (Sigma-Aldrich), Erastin (MedChem Express), ferrostatin-1 (Sigma-Aldrich), liproxstatin-1 (Sigma-Aldrich), NOX1/4 inhibitor (GKT137831, Cayman Chemical), Sulfasalazine (Sigma-Aldrich), Sorafenib (Sigma-Aldrich), RSL3 (MedChem Express), Glutathione (Sigma-Aldrich), N-acetyl-cysteine (Sigma-Aldrich), Ciclopirox olamine (CPX, Sigma-Aldrich), LOXi (PD-146176, Santa Cruz Biotechnology), Trolox (Sigma-Aldrich), Prankulast (Biomol), tunicamycin (Sigma-Aldrich), Erastin (MedChemExpress), BSO (Santa Cruz Biotechnology), cisplatin (Santa Cruz Biotechnology), doxorubicin (Sigma-Aldrich), 2-ME (Sigma-Aldrich), PPG (Sigma-Aldrich),
Nutlin-3 (Sigma-Aldrich), vildagliptin (Sigma-Aldrich), Q-VD-OPh hydrate (Sigma-Aldrich), bafilomycin A (Sigma-Aldrich), nocodazole (Sigma-Aldrich), panobinostat (Sigma-Aldrich), doxorubicin (Sigma-Aldrich). AMF-26 was synthesized by Merck KGaA^{23 (link)}.

Alborzinia H., Ignashkova T.I., Dejure F.R., Gendarme M., Theobald J., Wölfl S., Lindemann R.K, & Reiling J.H. (2018). Golgi stress mediates redox imbalance and ferroptosis in human cells. Communications Biology, 1, 210.

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Inhibition of Arf1 GEFs and Dynein

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Brefeldin A (BFA), the primary targets of which are the high molecular weight Arf1 GEFs GBF1, BIG1 and BIG2, and the highly specific GBF1 inhibitor, golgicide A (GCA), were obtained from Sigma-Aldrich and used as described previously^{15 (link),27 (link),29 (link)}. The cytoplasmic dynein inhibitor, ciliobrevin D, was purchased from EMD Millipore (Billerica, MA, USA). Stock solutions were prepared in DMSO.

Walch L., Pellier E., Leng W., Lakisic G., Gautreau A., Contremoulins V., Verbavatz J.M, & Jackson C.L. (2018). GBF1 and Arf1 interact with Miro and regulate mitochondrial positioning within cells. Scientific Reports, 8, 17121.

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Signaling Pathway Modulation Protocol

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Brefeldin A and rapamycin was purchased from LC Labs. Golgicide A was from Sigma Aldrich. Gramicidin was from Molecular Probes. Nigericin, ruxolitinib, and tofacitinib were from Invivogen. Vinblastine was from Tocris. Human recombinant IFNβ and IFNγ were obtained from Millipore.

Gregory D.J., DeLoid G.M., Salmon S.L., Metzger D.W., Kramnik I, & Kobzik L. (2020). SON DNA-Binding Protein mediates macrophage autophagy and responses to intracellular infection. FEBS letters, 594(17), 2782-2799.

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DENV-2 Infection Inhibition Study

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Confluent A549 cells were infected with DENV-2 16681 using a multiplicity of infection of 10. After 1 hour, cells were washed with PBS and DMEM-F12 medium supplemented with 10% serum was added. At 8 hours post-infection fresh medium with 0, 0.05, 0.1 or 0,5 µg/ml of Brefeldin A (Sigma), or 50 µM of Golgicide A (Sigma) was added. The medium was supplemented with 100 µM of oleic acid (Sigma) complexed with BSA (60 (link)). For immunofluorescence the cells were fixed at 24 hours post-infection.

Iglesias N.G., Mondotte J.A., Byk L.A., De Maio F.A., Samsa M.M., Alvarez C, & Gamarnik A.V. (2015). Dengue Virus Uses a Non-Canonical Function of the Host GBF1-Arf-COPI System for Capsid Protein Accumulation on Lipid Droplets. Traffic (Copenhagen, Denmark), 16(9), 962-977.

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High-resolution Drug Assays in RPE1 Cells

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In drug assays for 60× resolution experiments, RPE1 cells were treated with NZ (0.1 µM) (Sigma-Aldrich, St.Louis, MO) in complete DMEM/F-12 medium at 37°C in 5% CO₂ for the duration of the experiment. Also 30 min before the experiment and before addition of the drug, cells were incubated with Hoechst 33,342 (1 µg/ml) dye (Thermo Fisher Scientiﬁc, Waltham, MA), to label cell nuclei. Then, the dye was washed with 1× PBS buﬀer (pH = 7.5) and a drug, diluted in complete medium, was added. Cells were imaged immediately after addition of a drug.
In drug assays for Cytonote 6 W experiments, RPE1 cells were treated with NZ (0.1 µM) (Sigma-Aldrich, St.Louis, MO), Golgicide A (0.7 µM, 7 µM, 10 µM, 14 µM, 35 µM and 70 µM) (Sigma-Aldrich, St.Louis, MO), or Taxol (Paclitaxel) (0.1 µM) (Sigma-Aldrich, St.Louis, MO) in complete DMEM/F-12 medium at 37°C in 5% CO₂ for the duration of the experiment.

Vaidžiulytė K., Macé A.S., Battistella A., Beng W., Schauer K, & Coppey M. (2022). Persistent cell migration emerges from a coupling between protrusion dynamics and polarized trafficking. eLife, 11, e69229.

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Fatty Acid-Mediated Autophagy Modulation

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Culture media and supplements for cell culture were purchased from Gibco-Life Technologies (Carlsbad, CA, USA) and plasticware from Greiner Bio-One (Monroe, CA, USA). All cells were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 units/ml penicillin G sodium and 100 μg/ml streptomycin sulfate at 37 °C under 5% CO₂. All FAs were purchased from Larodan (Malmö, Sweden); golgicide A, rapamycin, thapsgargin and tunicamycin were obtained from Sigma-Aldrich (St. Louis, MO, USA). Each fatty acid was dissolved at a concentration of 100 mM in an appropriate volume of 100% ethanol pre-warmed at 37 °C. The obtained solution was then used to treat the cells, with final concentrations ranging from 125 to 1000 μM. The SCREEN-WELL® Autophagy library (BML-2837) was purchased from Enzo Life Sciences (Farmingdale, NY, USA).

Sauvat A., Chen G., Müller K., Tong M., Aprahamian F., Durand S., Cerrato G., Bezu L., Leduc M., Franz J., Rockenfeller P., Sadoshima J., Madeo F., Kepp O, & Kroemer G. (2018). Trans-Fats Inhibit Autophagy Induced by Saturated Fatty Acids. eBioMedicine, 30, 261-272.

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