Figure 5.

ROS control LPS- and high salt-induced suppression of NFAT5 target genes. (a and b) ROS dependency of NFAT5 target gene expression induced by LPS or high salt. The mRNA expression of AR, SMIT, IL-6 and NOS2 was determined by real-time PCR analysis 4 h after stimulation with NaCl or LPS. The data are the mean±s.d. of three independent experiments. *P<0.05 versus NaCl (A) or LPS alone (B). (c and d) ROS inhibitors restore SMIT, IL-6 and MCP-1 expression after combined treatment with high salt and LPS. RAW 264.7 macrophages were pretreated with various concentrations of allopurinol (Allo), rotenone (Ro), or myxothiazol (MTZ) for 1 h and then stimulated with LPS (5 μg ml⁻¹) and NaCl (90 mℳ) for 4 h. IL-6 and MCP-1 concentrations in the culture supernatants were assessed by enzyme-linked immunosorbent assay. SMIT mRNA expression was determined by real-time PCR. *P<0.01. The degree of cell viability was assessed by an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and presented as the percentage of the cell viability of treated samples to the viability of control untreated cells.