Methods and reagents for the treatment of immunoinflammatory disorders
A drug and inflammatory technology, applied in the field of treatment of immune inflammatory diseases and drug testing, can solve problems such as adverse side effects and effect changes
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[0141] Example 1: Inhibition of parallel signaling pathways by amoxapine and paroxetine
[0142] Materials and methods
[0143] drug
[0144] Stock solutions of all drugs were prepared in DMSO except amoxapine which was prepared in 0.1 mM MES (2-(N-morpholinoethanesulfonic acid) (Sigma) buffer. Myristic acid-phorbol-acetate (PMA) (100 μg / ml) stock solution and ionomycin (5 mg / ml) in DMSO were diluted in culture medium to produce final concentrations of PMA (10 ng / ml, 16.2 nM) and ionomycin ( 750 μg / ml, 1 μM).
[0145] Cells and Cell Lines
[0146] Fresh buffy coat preparations prepared from donated human whole blood (Red Cross, Rhode Island) were used to isolate peripheral blood mononuclear cells (PBMCs) by Ficoll-Plaque (Pharmacia) layered centrifugation. T cells were purified from PBMCs using "Pan T cell Isolation Kit II-human", (Miltenenyibiotec, Germany). The lymphoid leukemia T cell line (CCRF-CEM) was obtained from American Type Cell Culture (ATCC). All cells were ...
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