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. 2014 Jul 15;12(7):4214–4230. doi: 10.3390/md12074214

Figure 2.

Figure 2

Effects of fucoxanthin treatment on the expression, nuclear translocation, and antioxidant response element (ARE) sequence-binding activity of Nrf2. (A) Nuclear extracts were prepared from HaCaT cells following treatment with 20 μM fucoxanthin for the indicated amount of time. Western blotting of the nuclear lysates was performed using Nrf2 and phospho Nrf2 antibodies. * and # indicates significantly different from Nrf2 and phospho Nrf2 of control, respectively (p < 0.05); (B) An anti-Nrf2 antibody and a FITC-conjugated secondary antibody were used to detect Nrf2 localization (green) by using confocal microscopy.DAPI staining indicates the locations of nuclei (blue). The merged images show the nuclear localization of Nrf2 protein; (C) Nuclear extracts were prepared from HaCaT cells treated with 20 μM fucoxanthin for 6 h. A ChIP assay was performed to assess binding of Nrf2 to the ARE in the promoters of the genes encoding GCLC and GSS; (D) Transcriptional activity of Nrf2 in HaCaT cells following treatment with 20 μM fucoxanthin for 6 h was assessed by using luciferase reporter assay. * Significantly different from control cells.