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. 2014 Aug 25;289(41):28660–28675. doi: 10.1074/jbc.M114.595496

FIGURE 5.

FIGURE 5.

Nrf2 plays a critical role for cell survival in the cadmium-transformed BEAS-2B cells. The basal expression level of Nrf2 was measured in the cadmium-transformed BEAS-2B cells and non-transformed BEAS-2B cells by Western blot analysis (A). The normal and transformed BEAS-2B cells were exposed to various concentrations of cadmium (0–10 μm) for 24 h (B) or various times (0–24 h) with 10 μm cadmium (C); then the levels of Nrf2 were detected. To diminish Nrf2 levels, cells were transfected with siRNA specific to Nrf2. After overnight transfection, cells were exposed to 10 μm cadmium for an additional 24 h. The expression level of Nrf2 was evaluated by Western blot (D), and an apoptosis assay was performed after staining with annexin V/PI and is represented in dot blots (E) and a graph (F). Finally, the total number of cell deaths was calculated by the early (annexin V+/PI−) and late apoptotic (annexin V+/PI+) cells and necrotic (annexin V−/PI+) cells (G). The results represent the mean ± S.E. (error bars) of three independent experiments. **, p < 0.01, significant difference between the experiments (ANOVA, Scheffe's test). GAPDH was used as loading control.