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. 2018 Nov 9;294(1):101–115. doi: 10.1074/jbc.RA118.002896

Figure 6.

Figure 6.

Rnf183 enhancer region depends on NFAT5. A, effect of NFAT5 knockdown on hypertonicity-induced luciferase activities. Cells cotransfected with 3.5 kbp-Luc and siRNA (NFAT5 1 or NC) were treated with isotonic or 75 mm NaCl– or sucrose–supplemented medium for 24 h (n = 3). B, effect of rescue with FLAG-NFAT5 on luciferase activities. Cells pretransfected with siRNA (NFAT5 3 or NC) were cotransfected with 3.5 kbp-Luc and either empty or pFLAG-NFAT5 vectors after 12 h of knockdown. After 24 h of cotransfection, cells were treated with isotonic or 75 mm NaCl–supplemented medium for an additional 24 h (n = 4). C, effect of p38/MAPK inhibitor SB203580 on Rnf183 luciferase activities. Cells transfected with 3.5 kbp-Luc were treated with 75 mm NaCl for 24 h in the presence of the indicated concentrations of SB203580 (1 h of preincubation) (n = 3). D, effect of overexpression with FLAG-DN-NFAT5 and FLAG-NFAT5 on luciferase activities. Cells cotransfected with 3.5 kbp-Luc and either empty, pFLAG-DN-NFAT5, or pFLAG-NFAT5 vectors were analyzed after 48 h of transfection (n = 4). Data were analyzed by one-way ANOVA, followed by post hoc tests using t tests with Bonferroni correction. Values represent mean ± S.D. (error bars). *, p < 0.05; **, p < 0.01; ***, p < 0.001.