Figure 1. Oxidized extracellular E_h Cys/CySS increases pro-IL-1β in monocytes.

U937 cells were exposed to physiological (−80 mV) and oxidized (−46 mV) E_h Cys/CySS for 8 h and levels of the IL-1β precursor were determined by Western blot (A). Western blot analysis of the cell extract revealed increase in the 31 kDa precursor form of IL-1β at −46 mV compared to –80 mV. Quantitative analysis of the band densities of three separate experiments is shown as a bar graph. In (B) IL-1β levels determined by ELISA are expressed relative to protein concentration in the cell-supernatant. In (C), total RNA was extracted 4 h after treatment with redox media. Abundance of IL-1β mRNA was detected by real-time PCR and is normalized to β actin. In (D), U937 cells expressing the IL-1β-luciferase construct were exposed to given E_h for 12 h. Luciferase activity in cell-lysates is shown after normalization for total protein. Data are mean+SE of 3 replicates of a representative experiment repeated 3 times, * P<0.05 between −80 mV and −46 mV treatments.