Figure 2
CPUY192018 activated the Nrf2-ARE pathway in vitro.
(A) Effect of CPUY192018 on the induction of the Nrf2 protein expression. (B) Effect of CPUY192018 on the nuclear translocation of the Nrf2 protein. At various time poits after the treatment with CPUY192018 (10 μM), nuclear and cytoplasmic cell extracts were prepared from the NCM460 cells and subjected to western blot analysis. Histone and β-actin served as markers for nuclear and cytosolic Nrf2 proteins, respectively. Densitometric analysis was performed to determine the relative ratios of the protein in each fraction. The data were normalized to the β-actin expression and are expressed as the means + SEM of three individual experiments. The data were analyzed using Image J 1.44p. (C) Immunofluorescence staining of Nrf2 at the indicated times in the NCM460 cells treated with 10 μM CPUY192018. Nrf2 and the nuclei were labeled with FITC and DAPI, respectively. The bars indicate the magnification (10 μm). (D) ARE induction by CPUY192018 and t-BHQ in the HepG2−ARE−C8 cells. The cells were exposed to the compounds or DMSO for 12 h. The activities are shown as the ratio to the DMSO control. The values shown are the means ± SEM (n = 3 independent observations).
