Dexamethasone (dex)

Manufactured by Selleck Chemicals

Sourced in United States, China

Dexamethasone is a synthetic glucocorticoid medication used as a lab reagent. It is a potent anti-inflammatory and immunosuppressant agent. Dexamethasone is commonly used in research applications to study the effects of glucocorticoids on various biological processes.

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53 protocols using dexamethasone (dex)

Murine Xenograft Model of BCP-ALL

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Then, 1 × 10⁶ spleen cells from MH/NRAS^G12D BCP-ALL mice of secondary transplantations were transplanted into sublethally irradiated (450 cGy) recipient mice. The MH/NRAS^G12D tertiary recipient mice were subjected to the treatment of solvent (as control), 2.5 mg/kg panobinostat (Selleck, S1030), 0.15 mg/kg vincristine (VCR) (Selleck, S1241) plus 1 mg/kg dexamethasone (DEX) (Selleck, S1322), and panobinostat in combination with VCR + DEX, intraperitoneally for 4 cycles of 5-days-on/2-days-off after transplantation. panobinostat was diluted in 2% dimethyl sulfoxide, 48% PEG 300, 2% Tween 80, and double-distilled water. VCR was diluted in double-distilled water. DEX was diluted in 5% dimethyl sulfoxide, 45% PEG 300, and double-distilled water.
More information concerning materials and methods used in this study are described in the supplemental Materials and Methods.

Zhang M., Zhang H., Li Z., Bai L., Wang Q., Li J., Jiang M., Xue Q., Cheng N., Zhang W., Mao D., Chen Z., Huang J., Meng G., Chen Z, & Chen S.J. (2022). Functional, structural, and molecular characterizations of the leukemogenic driver MEF2D-HNRNPUL1 fusion. Blood, 140(12), 1390-1407.

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Differentiation of Adipose-Derived Mesenchymal Stem Cells

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Osteogenic differentiation: The ADMSCs at passage 4 were cultured in a 12-well plate with ADMSCs medium. When the cells growth reached 60% confluence, the ADMSCs were replaced with osteogenic differentiation medium (MEM medium containing 10% FBS, 100 nM dexamethasone (Selleck), 30 μg/ml ascorbic acid (sigma) and 10 mM sodium ß-glycerophosphate (Solarbio)) and differentiated for another 14 days. After that, the cells were fixed with 4% paraformaldehyde, washed with PBS, and incubated with alizarin red staining solution for 30 min. Finally, cells are used for photography after removing alizarin red staining solution and washing with PBS for three times.
Adipogenic differentiation: The ADMSCs were cultured similarly as above, then replaced with adipogenic differentiation medium (MEM medium containing 10% FBS, 5% Insulin-Transferrin-Selenium-Ethanolamine (ITS-X) (Selleck), 1 μM dexamethasone (Selleck) and 0.5 mM 3-Isobutyl-1-methylxanthine (IBMX) (Selleck)). After 14 days’ differentiation, the cells were fixed with 4% paraformaldehyde, washed with PBS, and incubated with oil red O staining solution (Solarbio) for 60 min. Finally, cells are used for photography after removing staining solution and washing with PBS for three times.

Li W., Wang W., He X., Liao Z., Aierken A., Hua J., Wang Y., Lu D, & Zhang S. (2022). Rapid recovery of male cats with postrenal acute kidney injury by treating with allogeneic adipose mesenchymal stem cell-derived extracellular vesicles. Stem Cell Research & Therapy, 13, 379.

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Flavopiridol and HDAC Inhibitor Treatments

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Cells were grown for 24 h after they were seeded onto the cell culture plates. For the flavopiridol time course experiment, 0.9 μL of 1 mM flavopiridol (Selleck Chemicals) was added to each 6 well to attain a final concentration of 300 nM. For the HDAC inhibitor time course experiment, 1 μL of 1 mM of either abexinostat (Selleck Chemicals) and pracinostat (Selleck Chemicals) was added to each 96 well to attain a final concentration of 10 μM. Similarly, for the HDAC inhibitor and dexamethasone co-treatment experiment, we added 1 μL of 100 μM or 1 μL of 1 mM of either abexinostat or pracinostat to each 96 well to attain a final concentration of 1 and 10 μM, respectively. dexamethasone (Selleck Chemicals) was added at a concentration of 100 nM (1 μL of 10 μM) two hours before the end of the HDAC inhibitor treatment. DMSO (10%) was used as a vehicle for flavopiridol and HDAC inhibitor treatments and ethanol (10%) was used as a vehicle for dexamethasone treatment. The HDACi timecourse and HDACi/DEX co-treatment experiment was performed in duplicates, and the flavopiridol timecourse experiment was only performed once.

Kim H.J., Booth G., Saunders L., Srivatsan S., McFaline-Figueroa J.L, & Trapnell C. (2022). Nuclear oligo hashing improves differential analysis of single-cell RNA-seq. Nature Communications, 13, 2666.

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Co-culture of Thymocytes and Dendritic Cells

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Co-culture experiments were carried out using exECs or DCs and thymocytes harvested from mechanically dissociated thymus from untreated mice, or in the case of DC analysis whole thymus cultures were used. Harvested thymocytes were incubated with either 100 nM dexamethasone (D2915, Sigma Aldrich, Germany), or 20 μM z-VAD-FMK (2163, Tocris, UK) for 4 hours at 37°C prior to co-culture, washed twice with PBS, and resuspended in exEC media for co-culture (1 × 10⁶ cells / well). Cells were harvested 20 hours post co-culture and prepared for either qPCR analysis or flow cytometry analysis. Thymic DCs were isolated from untreated mice using CD11c UltraPure microbeads (130–108-338, Miltenyi Biotech, USA), on enzymatically digested thymuses. DCs were cultured in DMEM (11965, GIBCO), 10% FBS (SH30066.03, HyClone, GE Life Sciences), and 1% PenStrep (15240–062, Invitrogen). For TAM receptor inhibitor studies, exECs were treated with 25 μM RXDX-106 (CEP-40783, s8570, Selleck Chemicals) 30 minutes prior to incubation with dexamethasone treated or z-VAD-FMK treated thymocytes, and Bmp4 expression was determined by qPCR analysis 20 h post co-culture. HEK293 cells (ATCC, Manassas, VA) were cultured in DMEM (11965, GIBCO), 10% FBS (SH30066.03, HyClone, GE Life Sciences), 1% Glutamax (35050061, Life Technologies), and 1% PenStrep (15240–062, Invitrogen).

Kinsella S., Evandy C.A., Cooper K., Iovino L., deRoos P.C., Hopwo K.S., Granadier D.W., Smith C.W., Rafii S, & Dudakov J.A. (2021). Attenuation of apoptotic cell detection triggers thymic regeneration after damage. Cell reports, 37(1), 109789.

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Anticancer Drug Preparation Protocol

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Enzalutamide (MedChemExpress), R1881 (Steraloids Inc. Newport RI), the AKR1C3 inhibitor (AKRi) (3-(4-trifluoromethyl)phenylamino) benzoic acid, Calbiochem), indomethacin (Sigma-Aldrich), dexamethasone (Selleck Chemicals), docetaxel (MedChemExpress) and mifepristone (Selleck Chemicals) were dissolved in dimethyl sulfoxide (DMSO). All solvents used for the isolation were purchased from VWR International (Darmstadt, Germany).

Kafka M., Mayr F., Temml V., Möller G., Adamski J., Höfer J., Schwaiger S., Heidegger I., Matuszczak B., Schuster D., Klocker H., Bektic J., Stuppner H, & Eder I.E. (2020). Dual Inhibitory Action of a Novel AKR1C3 Inhibitor on Both Full-Length AR and the Variant AR-V7 in Enzalutamide Resistant Metastatic Castration Resistant Prostate Cancer. Cancers, 12(8), 2092.

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Drug Sensitivity and Synergy Profiling in ALL

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Drug sensitivity and drug synergy profiles were evaluated using ALL cells in co-culture with MSCs as described above. A Tecan D300 robot was used to dispense drugs at the indicated dilutions. ABT-263 (navitoclax) and dexamethasone (Selleckchem) concentrations for synergy were based on the half maximal inhibitory concentration (IC₅₀) of the different samples screened. Cell viability was evaluated after 72 h of drug treatments using CyQuant staining and imaging-based cell viability analysis on an ImageXpress microscope (Molecular Devices; ref. ^{15 (link)}). All the dose response curves and the IC₅₀ were represented and calculated using GraphPad Prism 8. The synergy was calculated using SynergyFinder tool^{21 (link),22 (link)}. The resulting ZIP score indicates synergism (ZIP score ≥ 1), additivity (ZIP score 0–1) or antagonism (ZIP score ≤ 0), and is here represented by a 3D graph^{21 (link),22 (link)}.

Mezzatesta C., Abduli L., Guinot A., Eckert C., Schewe D., Zaliova M., Vinti L., Marovca B., Tsai Y.C., Jenni S., Aguade-Gorgorio J., von Stackelberg A., Schrappe M., Locatelli F., Stanulla M., Cario G., Bourquin J.P, & Bornhauser B.C. (2020). Repurposing anthelmintic agents to eradicate resistant leukemia. Blood Cancer Journal, 10(6), 72.

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Isolation and Culture of Primary Hepatocytes

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Primary hepatocytes were isolated from chow diet–fed WT, Asgr1^+/, and Asgr1^–/– mice by 2-step collagenase (MilliporeSigma) perfusion digestion and low-speed centrifugation (50g, 4°C), then cultured in the collagen-coated dish with DMEM (high glucose, Hyclone) containing 10% FBS (Gibco), 1% NEAA, 1% ITS (Gibco), 1% penicillin and streptomycin (Life Technologies), and 0.1 μM dexamethasone (Selleck). All cells were serum-starved for 12 hours before harvest.

Xu Y., Tao J., Yu X., Wu Y., Chen Y., You K., Zhang J., Getachew A., Pan T., Zhuang Y., Yuan F., Yang F., Lin X, & Li Y.X. (2021). Hypomorphic ASGR1 modulates lipid homeostasis via INSIG1-mediated SREBP signaling suppression. JCI Insight, 6(19), e147038.

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Prostate Cancer Compound Evaluation

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Apalutamide, darolutamide, dexamethasone, enzalutamide, mifepristone, OTX015, and JQ1 were purchased from Selleckchem. R1881, dihydrotestosterone (DHT), and beta-estradiol were purchased from Sigma-Aldrich. BET protein degrader ZBC260 and AR protein degrader ARD-61 were described previously (28 (link), 29 (link)).

Qiao Y., Wang X.M., Mannan R., Pitchiaya S., Zhang Y., Wotring J.W., Xiao L., Robinson D.R., Wu Y.M., Tien J.C., Cao X., Simko S.A., Apel I.J., Bawa P., Kregel S., Narayanan S.P., Raskind G., Ellison S.J., Parolia A., Zelenka-Wang S., McMurry L., Su F., Wang R., Cheng Y., Delekta A.D., Mei Z., Pretto C.D., Wang S., Mehra R., Sexton J.Z, & Chinnaiyan A.M. (2020). Targeting transcriptional regulation of SARS-CoV-2 entry factors ACE2 and TMPRSS2. Proceedings of the National Academy of Sciences of the United States of America, 118(1), e2021450118.

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Deubiquitinase Inhibition in Multiple Myeloma

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MM cell lines (MM.1S, MM.1R, RPMI8226, INA6, Dox40, LR5, ARP1) and peripheral blood mononuclear cells from normal healthy donors were cultured in RPMI-1640 medium containing 10% fetal bovine serum and antibiotics. ANBL6 cells were kindly provided by Dr Robert Orlowski (Houston, TX, USA).^{45 (link)} Tumor cells, BMSCs and pDCs were cultured as described previously.^{28 (link),29 (link)} Helsinki protocol was followed for obtaining informed consent from all patients. Bortezomib, dexamethasone, lenalidomide or pomalidomide were obtained from Selleck Chemicals LLC (Houston, TX, USA), and OPA was obtained from Calbiochem (San Diego, CA, USA). Recombinant DUBs were obtained from Creative BioMart (Rpn11/ PSMD14, Shirley, NY, USA), Boston Biochem (USP1, USP7, Cambridge, MA, USA) and LifeSensors Inc. (USP2, USP4, USP5, USP8, USP20, Uch37, Malvern, PA, USA).

Song Y., Li S., Ray A., Das D., Qi J., Samur M., Tai Y.T., Munshi N., Carrasco R., Chauhan D, & Anderson K. (2017). Blockade of deubiquitylating enzyme Rpn11 triggers apoptosis in multiple myeloma cells and overcomes bortezomib resistance. Oncogene, 36(40), 5631-5638.

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Dexamethasone-Induced Thymus Apoptosis in Mice

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Female C57BL/6 N mice were purchased from Orient Bio (Seongnam, Korea). All mice were acclimatized for 2 weeks at the laboratory animal research facility (National Cancer Center, Korea). The mice were administered 25 mg/kg BMS794833 (Cayman, USA) or control (5% DMSO, 10% kolliphor in PBS) by intraperitoneal injection. After 30 min, 8 mg/kg dexamethasone (Selleckchem, USA) was administered to the mice by intraperitoneal injection to induce thymus apoptosis. The thymus was collected and dissociated into a single-cell suspension as previously described in refs. ^{4 (link),23}. The thymocytes were analyzed by annexin V and propidium iodide staining and flow cytometry at the Flow Cytometry Core Facility (National Cancer Center, Korea) using FACSVerse (BD Bioscience, USA) as described previously^{24 (link)}.

Bae S.H., Kim J.H., Park T.H., Lee K., Lee B.I, & Jang H. (2022). BMS794833 inhibits macrophage efferocytosis by directly binding to MERTK and inhibiting its activity. Experimental & Molecular Medicine, 54(9), 1450-1460.

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