For cytokine stimulation of Ba/F3 cells, cells were washed twice in RPMI 1640 and rested in RPMI-based medium for 12 hours before addition of 5 ng/mL IL-3. Duration of IL-3 stimulation was contingent upon the downstream assay, as indicated in the figure legends. For chalcone and other inhibitor treatments, Ba/F3 cells were incubated with compounds or DMSO (vehicle) for 30 minutes prior to cytokine stimulation. Ba/F3-1*6 and K562 cells were treated for 60–90 minutes with the various compounds or DMSO (vehicle), as indicated. With the exception of cell viability assays (see below), all experiments were performed in the presence of equal amounts of DMSO.
Culturing Cell Lines for Cytokine Stimulation
For cytokine stimulation of Ba/F3 cells, cells were washed twice in RPMI 1640 and rested in RPMI-based medium for 12 hours before addition of 5 ng/mL IL-3. Duration of IL-3 stimulation was contingent upon the downstream assay, as indicated in the figure legends. For chalcone and other inhibitor treatments, Ba/F3 cells were incubated with compounds or DMSO (vehicle) for 30 minutes prior to cytokine stimulation. Ba/F3-1*6 and K562 cells were treated for 60–90 minutes with the various compounds or DMSO (vehicle), as indicated. With the exception of cell viability assays (see below), all experiments were performed in the presence of equal amounts of DMSO.
Corresponding Organization :
Other organizations : University of Regensburg
Protocol cited in 4 other protocols
Variable analysis
- Chalcone and other inhibitor treatments
- Duration of IL-3 stimulation
- Cell viability
- Downstream assay outcomes
- RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum (FCS), penicillin/streptomycin
- Incubation at 37°C under 5% CO2 in a humidified incubator
- Equal amounts of DMSO in all experiments (except cell viability assays)
- K562 cells
- Ba/F3 cells
- Ba/F3-1*6 cell line
- DMSO (vehicle)
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