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Ruxolitinib

Manufactured by Axon Medchem
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Ruxolitinib is a laboratory reagent used for research purposes. It is a small molecule inhibitor that targets the Janus kinase (JAK) family of enzymes.

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Lab products found in correlation

Tgf β1, by Thermo Fisher Scientific (2 mentions) Celltiter 96 aqueous one, by Promega (1 mentions) Sp 100030, by Bio-Techne (1 mentions) Velcade, by Selleck Chemicals (1 mentions) Sh 4 54, by Selleck Chemicals (1 mentions) As1517499, by Axon Medchem (1 mentions) Tofacitinib, by Selleck Chemicals (1 mentions) Belinostat, by Selleck Chemicals (1 mentions) Romidepsin, by MedChemExpress (1 mentions) Trichostatin a, by Selleck Chemicals (1 mentions) Stattic, by Axon Medchem (1 mentions) 5 aza, by Merck Group (1 mentions) Vorinostat, by Selleck Chemicals (1 mentions) Ly294002, by Cayman Chemical (1 mentions) Blebbistatin, by Merck Group (1 mentions) Recombinant human lif, by Merck Group (1 mentions) Cyt387, by Selleck Chemicals (1 mentions) Y 27632, by Bio-Techne (1 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)

6 protocols using ruxolitinib

1

Evaluating JAK1/2 and IGF1R Inhibitors

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Patients were seen at the Oregon Health & Science University Center for Hematologic Malignancies (patients #1 to #3, #5 and #6) or at the Hematology and Hemotherapy Center of the University of Campinas (patients #4 and #7). Following informed consent, peripheral blood was collected and mononuclear cells were isolated by Ficoll-gradient centrifugation. Cells were resuspended in RPMI 1640 medium supplemented with 10% FBS, L-glutamine, and penicillin/streptomycin and plated in 384-well plates (9000 cells/well) in triplicate in the presence of the JAK1/2 inhibitor ruxolitinib (0-4000nM), the IRS/IGF1R inhibitor NT157 (0-4000nM; Axon MedChem, USA), or both inhibitors in combination at the indicated concentrations. Plates were cultured at 37°C for 72h, and absorbance was measured by a methanethiosulfonate (MTS)-based assay, according to the manufacturer's protocol (CellTiter 96 AQueous One; Promega) for patients #1 to #3, #5 and #6, or by methylthiazole tetrazolium (MTT) assay for patients #4 and #7. Data were normalized to the mean percent of the untreated control wells.
de Melo Campos P., Machado-Neto J.A., Eide C.A., Savage S.L., Scopim-Ribeiro R., da Silva Souza Duarte A., Favaro P., Lorand-Metze I., Costa F.F., Tognon C.E., Druker B.J., Saad S.T, & Traina F. (2016). IRS2 silencing increases apoptosis and potentiates the effects of ruxolitinib in JAK2V617F-positive myeloproliferative neoplasms. Oncotarget, 7(6), 6948-6959.
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2

CHOP Cytotoxicity in Farage Cells

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CHOP consisted of four drugs (cyclophosphamide, vincristine, adriamycin, and prednisone) in a ratio of 80/5.5/0.16/11.1, respectively [45 (link)]. All four drugs were purchased from Selleckchem (Houston, TX, USA). Farage cells were treated with various concentrations of CHOP for 2 days or 3 days. Other drugs and their sources were as follows: Velcade (Selleckchem), ruxolitinib (Axon Medchem, Groningen, Netherlands), lenalidomide (Axon Medchem), SP 100030 (Tocris Bioscience, Bristol, United Kingdom), pimozide (Calbiochem, Darmstadt, Germany), AS 1517499 (Axon Medchem) and SH-4-54 (Selleckchem). The determination of IC50 and generation of the graphs were accomplished using GraphPad software.
Yoon H, & Ko Y.H. (2017). LMP1+SLAMF1high cells are associated with drug resistance in Epstein-Barr virus-positive Farage cells. Oncotarget, 8(15), 24621-24634.
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3

Inhibitor Screening for Epigenetic Targets

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Chidamide was provided by Shenzhen Chipscreen Biosciences Ltd. SAHA was obtained from Sigma–Aldrich. Trichostatin A (TSA), belinostat and tofacitinib were purchased from Selleckchem. Romidepsin was obtained from MedChemExpress. Ruxolitinib and Stattic were purchased from Axon Medchem and Millipore, respectively.
Chen J., Zuo Z., Gao Y., Yao X., Guan P., Wang Y., Li Z., Liu Z., Hong J.H., Deng P., Chan J.Y., Cheah D.M., Lim J., Chai K.X., Chia B.K., Pang J.W., Koh J., Huang D., He H., Sun Y., Liu L., Liu S., Huang Y., Wang X., You H., Saraf S.A., Grigoropoulos N.F., Li X., Bei J., Kang T., Lim S.T., Teh B.T., Huang H., Ong C.K, & Tan J. (2023). Aberrant JAK-STAT signaling-mediated chromatin remodeling impairs the sensitivity of NK/T-cell lymphoma to chidamide. Clinical Epigenetics, 15, 19.
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4

Cytokine Signaling Inhibitor Screening

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TGFβ1 was purchased from Peprotech (#100-21, Peprotech, Rocky Hill, NJ) and was used at 2 ng ml−1; recombinant human LIF was purchased from Millipore (#LIF1005,Millipore, Billerica, MA), and was used at a concentration of 2 ng ml−1. LIF neutralizing antibody (AB-250-NA, R&D, Minneapolis, MN) was used at 10 μg ml−1. Neutralizing antibodies were incubated for 1 h with CM before experiments. The following inhibitors were used in this study: Pyridone 6 (#42009, Calbiochem, Los Angeles, CA) was used at 5 μM, Ruxolitinib (#1598, Axon medchem, Groningen, The Netherlands) at 10 μM, 5′-Aza (#A2385, Sigma, Saint Louis, USA) was used daily at 2.5 μM, PTP inh 1 (#540200, Millipore, Billerica, MA) at 10 μM, Sodium Orthovanadate (#567540, Millipore, Billerica, MA) at 1 mM, C646 (#SML002, Sigma, Saint Louis, USA) and CTPB (#sc- 202558, Santa Cruz, Biotechnology, Santa Cruz, CA) at 25 μM. A list of the concentrations used for the inhibitors/activators in Fig. 1d,e is provided in Supplementary Table 1.
Albrengues J., Bertero T., Grasset E., Bonan S., Maiel M., Bourget I., Philippe C., Herraiz Serrano C., Benamar S., Croce O., Sanz-Moreno V., Meneguzzi G., Feral C.C., Cristofari G, & Gaggioli C. (2015). Epigenetic switch drives the conversion of fibroblasts into proinvasive cancer-associated fibroblasts. Nature Communications, 6, 10204.
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5

Evaluating MPN Cell Responses to Stromal Interactions

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HS-5 and KM-102 cell lines were cultured to 70% confluence and the MPN cells added to the stromal layer of HS-5 (+ HS-5) or KM-102 (+ KM-102) at 0.1x106 cells/ml in the appropriate culture medium, either directly (for cell to cell contact) or indirectly (separated by a 0.4-mm-thick micropore membranes +HS-5 TW). In addition, MPN cells were incubated without any stromal support (no stroma) or with the HS-5 conditioned media (+ CM), diluted 50% in the respective culture media. Vorinostat (Selleckchem), Ruxolitinib (Axon Medchem), SP600125 (JNK inhibitor) (Selleckchem) and LY294002 (PI3K inhibitor) (Cayman Chemicals) were added to the co-cultures once the MPN cells adhered to the BM stroma. At the indicated time points the cells were harvested and assessed as described below for viability, gene expression and immunoblotting.
Cardoso B.A., Belo H., Barata J.T, & Almeida A.M. (2015). The Bone Marrow-Mediated Protection of Myeloproliferative Neoplastic Cells to Vorinostat and Ruxolitinib Relies on the Activation of JNK and PI3K Signalling Pathways. PLoS ONE, 10(12), e0143897.
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6

Cytokine Signaling Pathway Modulation

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TGFβ1 was purchased from Peprotech (#100-21, Peprotech, Rocky Hill, NJ) and was used at 2 ng/ml; recombinant human GCSF (#300-23) and IL-6 (#200-06) were purchased from Peprotech and were used at 10ng/mL, recombinant human LIF was purchased from Millipore (#LIF1005, Millipore, Billerica, MA), and was used at a concentration of 2 ng/ml. Recombinant TNF alpha was produced in E Coli and purified under native conditions using an N-terminal 6-his tag. ICAM-1 neutralizing antibody (#BBA3, R&D, Minneapolis, MN) was used at 10 μg/ml. The following inhibitors were used in this study: Ruxolitinib (#1598, Axon medchem, Groningen, The netherlands) at 10μM, CYT387 (#S2219, Selleckchem, Huston, TX) at 10μM Y27632 (#1254, Tocris bioscience, Ellisville, MO) at 10μM, Blebbistatin (#B0560, Sigma, Saint Louis, MO) at 10μM, SU6656 (#572635, CalbioChem, Los Angeles, CA) at 10μM and Verteprofin (#SML0534, Sigma, Saint Louis, MO) at 4μg/mL.
Bonan S., Albrengues J., Grasset E., Kuzet S.E., Nottet N., Bourget I., Bertero T., Mari B., Meneguzzi G, & Gaggioli C. (2016). Membrane-bound ICAM-1 contributes to the onset of proinvasive tumor stroma by controlling acto-myosin contractility in carcinoma-associated fibroblasts. Oncotarget, 8(1), 1304-1320.
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