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Everolimus

Manufactured by Cell Signaling Technology
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Everolimus is a small-molecule inhibitor that targets the mammalian target of rapamycin (mTOR) pathway. It functions by binding to the intracellular protein FKBP-12, forming a complex that inhibits the activity of mTOR, a key regulator of cell growth, proliferation, and survival.

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Lab products found in correlation

Wst 1 proliferation reagent, by Roche (2 mentions) Siomycin a, by Merck Group (2 mentions) Cell culture reagents, by Corning (1 mentions) One glo luciferase assay, by Promega (1 mentions) Torin 1, by Cell Signaling Technology (1 mentions) Celltiter fluor cell viability assay, by Promega (1 mentions) Pluronic p103, by BASF (1 mentions) Muramyl dipeptide mdp, by InvivoGen (1 mentions) Chloroquine diphosphate, by MP Biomedicals (1 mentions) Triton x 100, by Thermo Fisher Scientific (1 mentions) Pluronic f127, by BASF (1 mentions) Pp242, by Abcam (1 mentions) Pepstatin, by Merck Group (1 mentions) Rapamycin, by Merck Group (1 mentions) Torin1, by Bio-Techne (1 mentions) Acridine orange, by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Thymidine, by Merck Group (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Propidium iodide, by Merck Group (1 mentions)

5 protocols using everolimus

1

Lipid-based Delivery of Firefly Luciferase mRNA

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Guanabenz acetate, heparin sodium salt, ethanol, Synperonic PE/P84 (Pluronics® P84) were purchased from Sigma-Aldrich. Bioactive Lipid Library II in addition to compounds L1–L12 was obtained from Cayman Chemicals (Details in Supplementary Excel File). All leukotriene inhibitors were purchased from Cayman Chemicals. Chloroquine diphosphate was purchased from MP Biomedicals. Everolimus and Torin 1 were purchased from Cell Signaling Technologies. Pluronic® F127 and Pluronic® P103 were purchased from BASF. Muramyl dipeptide (MDP) was purchased from InvivoGen. Triton X-100 was purchased from Acros Organics. Quanti-iT RiboGreen RNA reagent and rRNA standards were purchased from Life Technologies. CellTiter Fluor Cell Viability Assay and One-Glo Luciferase Assay was purchased from Promega. Firefly luciferase mRNA (FLuc mRNA, Moderna) and lipid formulation (ionizable lipid: structural lipid: cholesterol: PEG-lipid) were obtained from Moderna Therapeutics. All cell culture reagents were purchased from Corning.
Patel S., Ashwanikumar N., Robinson E., DuRoss A., Sun C., Murphy-Benenato K.E., Mihai C., Almarsson Ö, & Sahay G. (2017). Boosting intracellular delivery of lipid nanoparticle-encapsulated messenger RNA. Nano letters, 17(9), 5711-5718.
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2

Autophagy Modulation with Small Molecules

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In this study, drugs and reagents included Rapamycin, 10 μg/mL E64d, 10 μg/mL pepstatin, Acridine Orange (AO), and dimethylsulfoxide (DMSO) (all from Sigma-Aldrich, St. Louis, MO, USA), Torin 1 (Tocris, Bristol, UK), pp242 (Abcam, Cambridge, MA, USA), everolimus (Cell Signaling Technology, Danvers, MA, USA), and FK-506 (tacrolimus) and pimecrolimus (both from Santa Cruz, Dallas, TX, USA). The control cells (the cells without drugs treatment or UVB radiation were named as nontreatment (NT)) were treated with 0.1% DMSO, which was used as the solvent for Rapamycin, E64d, pepstatin, Torin 1, pp242, everolimus, FK-506 (tacrolimus), and pimecrolimus. The DMSO solvent in our study was not beyond 0.1% [35 (link)].
Xu S., Li L., Li M., Zhang M., Ju M., Chen X, & Gu H. (2017). Impact on Autophagy and Ultraviolet B Induced Responses of Treatment with the MTOR Inhibitors Rapamycin, Everolimus, Torin 1, and pp242 in Human Keratinocytes. Oxidative Medicine and Cellular Longevity, 2017, 5930639.
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3

Cell Cycle Synchronization and Cell Proliferation Assays

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For cell cycle synchronization, cells were incubated in 10μM thymidine (Sigma-Aldrich, St. Louis, MO, USA; in culture media) for 72 hours or starved 24 hours in culture medium containing 0.01% FBS.
Siomycin A was obtained from Sigma-Aldrich and dissolved in DMSO. Everolimus as control therapy was obtained from Cell Signaling Technology. For proliferation and cytotoxicity assays, 10000-20000 cells per well were used in quintuplets in 96 well plates and treated 4 and 24h for LDH and 24, 48 and 72h for WST-assay with different concentrations and controls. WST-1 proliferation reagent and LDH cytotoxicity detection reagent (Roche; Basel, Switzerland) were applied according to the manufactures' instructions. In the LDH assay, 3% FBS was used instead of 1%, as recommended in the instructions. Cell density was colorimetrically quantified with a multi-well spectrophotometer (TECAN sunrise™).
Briest F., Berg E., Grass I., Freitag H., Kaemmerer D., Lewens F., Christen F., Arsenic R., Altendorf-Hofmann A., Kunze A., Sänger J., Knösel T., Siegmund B., Hummel M, & Grabowski P. (2015). FOXM1: A novel drug target in gastroenteropancreatic neuroendocrine tumors. Oncotarget, 6(10), 8185-8199.
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4

Evaluating AdipoRon and Everolimus Effects

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Bovine serum albumin (BSA) (Microtech, #B2518), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (Sigma Life Science), propidium iodide (PI) (Sigma Life Science, #P4864), AdipoRon (Focus Bioscience, St Lucia, QLD, Australia), and everolimus (Cell Signaling Technology, #12017).
Sapio L., Nigro E., Ragone A., Salzillo A., Illiano M., Spina A., Polito R., Daniele A, & Naviglio S. (2020). AdipoRon Affects Cell Cycle Progression and Inhibits Proliferation in Human Osteosarcoma Cells. Journal of Oncology, 2020, 7262479.
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5

Proliferation Assay for Nutlin-3, Everolimus, and Siomycin A

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Nutlin-3 (CAS No. 890090-75-2) was obtained from Santa Cruz Biotechnologies; Nutlin-3a (CAS No. 675576-98-4) was obtained from Selleck Chemicals, everolimus from Cell Signaling, and siomycin A from Sigma Aldrich. Substances were dissolved in DMSO. WST-1 proliferation assays were performed according to the manufacturer's instructions in quintuplets after treatment with decreasing concentrations of the substances. The WST-1 proliferation reagent was obtained from Roche, Basel, Switzerland. Cell density was colorimetrically quantified with a multiwell spectrophotometer (TECAN sunrise TM ) and analyzed with MS Excel 2013 and Prism 6.
Briest F., Grass I., Sedding D., Möbs M., Christen F., Benecke J., Fuchs K., Mende S., Kaemmerer D., Sänger J., Kunze A., Geisler C., Freitag H., Lewens F., Worpenberg L., Iwaszkiewicz S., Siegmund B., Walther W., Hummel M, & Grabowski P. (2018). Mechanisms of Targeting the MDM2-p53-FOXM1 Axis in Well-Differentiated Intestinal Neuroendocrine Tumors. Neuroendocrinology, 107(1).
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