Fig. 3.

A: LPS decreases expression of a chimeric zinc-sensitive reporter. SPAECs were transiently cotransfected with pLuc-MCS/4MREa (0.6 μg) and pSVβ-galactosidase (0.15 μg). 24 h after transfection, cells were treated with or without LPS (100 ng/ml) for 4 h in the presence of serum (10%) and subsequently assayed for luciferase and β-galactosidase (β-Gal) activity 4 h posttreatment. A significant decrease in luciferase activity was observed in LPS-stimulated cells with viability greater than 92%. Luciferase activity is expressed as a ratio of firefly luciferase (pLucMRE) activity to β-galactosidase activity. Data represent means ± SE of luciferase activity of samples measured in triplicate for 9 independent experiments (*P < 0.05; independent, two-tailed t-test). B: cell viability was assessed by Alamar Blue. Alamar Blue fluorescence was measured as an index of viability 4 h post-LPS treatment. Data represent Alamar Blue fluorescence (% of control) measured in triplicate.