(a)
(b)
Figure 5: BKFE activates the transcription factor Nrf2 in SV40 MES13 cells. (a) SV40 MES13 cells were treated with 0, 1, 5, or 20 μg/ml BKFE for 18 h. Cytoplasmic and nuclear Nrf2 proteins were extracted and analyzed by western blotting. Lamin B and β-actin were used as loading controls for the nuclear and cytosolic fractions, respectively. The relative expression of Nrf2 was quantified using the ImageJ software. (b) Cells were seeded on coverslips and treated with 0, 1, 5, or 20 μg/ml BKFE for 18 h. Immunocytochemistry was performed using an anti-Nrf2 antibody and FITC-conjugated secondary antibody. Cell nuclei were counterstained with DAPI (blue). Boxed areas are enlarged. Magnifications, ×200 and ×850 (boxed areas). The relative fluorescence expression of Nrf2 was quantified using the ImageJ software. p<0.05, p<0.01, and p<0.001.