Cardiotrophin-1 Induces Tumor Necrosis Factor Synthesis in Human Peripheral Blood Mononuclear Cells
Figure 6
Effect of parthenolide on CT-1 induced α expression. (a) Human PBMCs were incubated with CT-1 (50 ng/ml) for 1 hour in the presence of parthenolide. Afterwards cells were lysed and α mRNA expression was determined by real-time-PCR. All α mRNA expression data were normalized to GAPDH. , data are expressed as meanSEM. compared to unstimulated cells. (b) Human PBMCs were incubated with CT-1 (50 ng/ml) for 3 hours in the presence of parthenolide. Afterwards α protein concentration in the supernatant was determined by ELISA. , data are expressed as meanSEM and normalized to unstimulated cells. compared to unstimulated cells. significant compared to unstimulated cells. (c) Analysis of intracellular production using immunofluorescent flow cytometry. Human blood was incubated with 50 ng/ml CT-1 for 6 hours in the presence of brefeldin A and parthenolide. Afterwards erythrocytes were lysed and cells were stained with a monoclonal antibody against CD14 FITC-conjugated and against α PE-conjugated. Monocytes were gated and results are expressed normalised to unstimlulated monocytes. n=6, data are expressed as meanSEM. compared to unstimulated cells, compared to cells stimulated with 50 ng/ml CT-1.
