Figure 3. Effects of ERRα silencing on the SCT promoter in response to hyperosmotic shock.

(A) Cells were treated with various saline concentrations (25, 50, 100 mM) for different times (2, 4, 8 h) or ANGII (10⁻¹¹ to 10⁻⁷ M) for 8 h, and RNA was extracted afterwards. The mRNA levels of mouse ERRα were normalized with mouse GAPDH levels. Data represent the mean ± SEM of three experiments performed in duplicates.*p<0.05; **p<0.001, compared with the respective control. (B) Western blot analysis of ERRα protein in N-42 cells after saline (upper) and ANGII (10⁻¹¹ to 10⁻⁹ M). (C) Effect of saline and ANGII treatment on ERE-half site mutants. Cells were transfected with mSCTP, M2 or M3 (2.0 µg) for 2 d and treated with saline (100 mM) for different times or ANGII (10⁻⁸ M) for 8 h. Data represent the mean ± SEM of three experiments performed in triplicates. ^*p<0.05, compared with the control promoter without treatment. (D) N-42 cells were transfected with mSCTP (2.0 µg) and either pSilencer (Control) (2.0 µg) or pSi-mERRα-S1 (2.0 µg). After 2 d, cells were subject to treatement with saline-added medium of different concentrations (25, 50, 100 mM) or ANGII (10⁻¹⁰ to 10⁻⁷ M) for 8 h. Data represent the mean ± SEM of three experiments performed in triplicates.*p<0.05; **p<0.001, compared with the control transfected with pSilencer without treatment. ^#p<0.05, compared with the control transfected with pSi-mERRα-S1 without treatment.