Ba/F3 or Ba/F3-gp130 cells were retrovirally transduced with constructs for mbHIL-6, IL-11R, mbHIL-11, and gp130 variants as described (37). Transduced cells were grown in standard DMEM supplemented with HIL-6 (10 ng/ml) or IL-3 (10 ng/ml). Selection of transduced Ba/F3 cells was performed with puromycin (1.5 μg/ml) or hygromycin B (1 mg/ml) (Carl Roth GmbH) or both. HEK 293 cells were grown in standard DMEM. Cells (4 × 105) were transiently transfected with Turbofect and 1 μg of plasmid DNA for sgp130Fc and 100 ng of plasmid DNA for HIL-6Fc. Six hours after transfection, the medium was exchanged to standard DMEM without transfection reagent. Cells were harvested 48 hours after stimulation for 30 min as indicated.

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