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. 2017 Mar 31;18:261–273. doi: 10.1016/j.ebiom.2017.03.039

Fig. 4.

Fig. 4

KRN2 inhibits the expression of NFAT5 and its target genes in primary macrophages. (a) Murine peritoneal macrophages were incubated with LPS (1 μg/ml) for 24 h in the absence or presence of KRN2 (0.8 μM). NFAT5 expression levels were determined by western blot analysis. Data are the representative of three independent experiments. (b–c) Murine peritoneal macrophages were stimulated with LPS (10 and 100 ng/ml) (b) or NaCl (45 mM) (c) for 6 h in the absence or presence of KRN2 (0.8 μM). The mRNA expression levels of Nfat5, Il6, Tnf, Ar and Smit were assessed by real-time PCR. Data are the mean ± SD of three independent experiments in duplicates. *P < 0.05 and †P < 0.05 versus LPS alone. (d) Murine splenocytes were incubated with LPS (1 μg/ml) for 24 h in the absence or presence of KRN2 (0.8 μM). The expression levels of NFAT5 and iNOS were determined by western blot analysis. Data are the representative of three independent experiments with similar results. (e) Murine splenocytes were incubated in the same condition as described in (d). The concentration of IL-6 and TNF-α in the culture supernatants were measured by ELISA. Data shows the mean ± SD of three independent experiments. *P < 0.05, **P < 0.005 versus only LPS-stimulated cells.