Figure 3.

Effects of tolvaptan on angiotensin II (AngII)‐induced ERK and eIF2α phosphorylation and DDIT3 protein expression in H295R cells. ERK phosphorylation (a). Cells were pretreated with tolvaptan at the indicated doses for 30 min and then stimulated with 100 nmol·L⁻¹ AngII for 5 min. The phosphorylation level of ERK was analysed by Western blotting. Upper, representative immunoblots; lower, relative quantification of protein expression. (b) Effect of tolvaptan on DDIT3 protein expression levels. Cells were pretreated with tolvaptan at the indicated doses for 30 min and then stimulated with 100 nmol·L⁻¹ AngII for 24 hr. DDIT3 protein level was analysed by Western blotting. Upper, representative immunoblots; lower, relative quantification of protein expression. eIF2α phosphorylation (c). Cells were pretreated with tolvaptan at the indicated doses for 30 min and then stimulated with 100 nmol·L⁻¹ AngII for 24 hr. The phosphorylation level of eIF2α was analysed by Western blotting. Upper, representative immunoblots; lower, relative quantification of protein expression. Results are shown as mean ± SD (n = 5 separate experiments) and expressed as fold over control. ^† P ˂ 0.05 versus Cont; *P ˂ 0.05 versus AngII