FIG. 2.

SspH1 inhibits NF-κB-dependent reporter gene expression. CHO-K1 cells were transiently transfected with the indicated constructs: (A) a plasmid in which expression of firefly luciferase is under the control of the ELAM-1 promoter, a construct in which Renilla luciferase is expressed from the herpes simplex virus thymidine kinase promoter, vectors expressing mouse CD14 and the tetracycline transctivator, plus an empty, SspH1-expressing, or SspH1-nonexpressing vector (Reverse sspH1); (B and C) a plasmid in which expression of firefly luciferase is under the control of either ELAM-1 (B) or the Rous sarcoma virus promoter (C), a construct in which Renilla luciferase is expressed from the herpes simplex virus thymidine kinase promoter, vectors expressing mouse CD14 and the tetracycline transctivator, plus increasing concentrations of SspH1 expression vector (0, 2.5, 5, and 7.5 μg), while the amount of introduced DNA was kept constant by the addition of empty vector. After 24 h, the cells were incubated with E. coli LPS (solid bars) or fresh medium (open bars) for 5 h, and luciferase activity was measured. Firefly luciferase activity was normalized to the activity of Renilla luciferase. Data are represented as the mean ± standard deviation of triplicate samples.