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2 protocols using ro 61 8048

1

Investigating MM-pDC Interactions and Immunotherapies

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MM cells were cultured in 10% FBS plus RPMI-1640 medium supplemented with antibiotics. MM–pDCs were cocultured either in DCP-MM medium (Mattek Corp., Ashland, MA) or complete RPMI-1640 medium supplemented with IL-3 (Peprotech Inc., Rocky Hill, NJ, USA). CD3-PE/FITC/APC; CD4-FITC/PE or APC-Cy7; CD8-APC/FITC, CD56-PE; CD123-PE/PE-Cy5/FITC; and CD138-FITC/PE/APC were obtained from BD Biosciences (San Jose, CA). BDCA-2-FITC and CD11c-APC were obtained from Miltenyi Biotec (Auburn, CA); CD303-; CD304-; CD107a; and PD-L1-BV421 were purchased from Biolegend. All immunomagnetic separation kits were purchased from Miltenyi Biotec. The CellTrace Violet and CellTracker Green flow assay kits were obtained from Life Technologies (USA). Functional-grade PD-L1 blocking antibody (antihuman PD-L1, clone MIH1) was obtained from eBiosciences [4 (link)]. Ro 61–8048 [13 (link)] and INCB 024360 were purchased from Selleck Chemicals. WST-1 Cell Proliferation Reagent was purchased from Clontech Laboratories, Inc. (USA).
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2

Examining KMO Role in Blast Injury

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Mice were treated daily with Ro-61-8048 (Selleckchem, Houston, TX, USA; 10 mg/kg/day, oral) or vehicle (2% DMSO, 30% polyethylene glycol, 5% Tween, in water) for 3 days prior to blast injury, and continuously for the duration of the study. Mice exposed to blast to examine the role of KMO received one single 20-PSI injury. Control mice were exposed to sham injury.
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