Effect of ZER on ARE promoter activation and subsequent expression of HO-1 and γ -GCLC proteins in HSF cells. (a) Cells were treated with ZER (8 μ M for 2 h) and subcellular localization of Nrf2 was determined using immunostaining method. (b) HSF cells were cotransfected with pGL3-ARE and treated with various concentrations of ZER (2-8 μ M for 2 h) to measure the percentage of ARE promoter activity. Data was presented as fold over increase in the percentage of ARE promoter activity. (c, d) The effect of ZER treatment (8 μ M) on the expression of total Nrf2 and antioxidant proteins (HO-1 and γ -GCLC) at different time points (0, 1, 2, 4, 8, 12, 16, or 24 h) was measured using western blot method against β -actin as an internal control. Results from three or more experiments were presented as mean ± SD, and the statistical significance was considered as ∗ p
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