Cell viability and IL-32 expression in IFNγ and H
+ IFNγ stimulated HBE cells. HBE cells were incubated with 10 ng/ml IFNγ or 250 μM H2O2 and 10 ng/ml IFNγ for 48 hours. The viability of the cells was examined using MTT assay (A). HBE cells were incubated with or without 2 hour-H2O2 pretreatment and then with or without IFNγ stimulation for 0, 4, 8, and 24 hours. The expression levels of IL-32 mRNA adjusted with expression levels in non-stimulated cells in each time point were determined by quantitative real-time PCR. HBE cells stimulated with H2O2, with IFNγ, and with IFNγ and H2O2, were presented in square plots, triangle plots, and inverted-triangle plots, respectively in graph (B). The bars show the means ± SE from data performed on 3 different individuals. *p < 0.01 significantly different. (C) After 48 hours of IFNγ treatment 20 μg of cell lysates were subjected to Western blotting. The 22 kDa band represents IL-32β/δ protein, and the 26 kDa band represents IL-32γ protein. The results shown are representative of 3 independent experiments.