Bilirubin Oxidation End Products (BOXes) Induce Neuronal Oxidative Stress Involving the Nrf2 Pathway

Figure 6

BOX A and BOX B induce the nuclear accumulation of Nrf2 and promote the antioxidant responsive gene expression on the protein level. (a) Representative immunofluorescent staining of Nrf2 distribution and nuclear accumulation in SH-SY5Y cells. The cell treatment with BOX A or BOX B (30 μM) for 24 h was stained with anti-Nrf2 antibody (red) and counterstained with Hoechst33342 (blue). Bar 20 μm. (b) Intensity profile of different fluorescent staining as the yellow dashed line with arrow shown in (a) in pixel distance (left, Mock; middle, BOX A; right, BOX B). The intensity profile of the line was measured by profile of the ZEN 2.6 lite software (Carl-Zeiss). Red, Nrf2 staining; blue, Hoechst33342; green, background noise. (c) Western blot showing the protein expression of Nrf2 and downstream target gene expression in 11 DIV cultured PCNs treated with BOX A or BOX B (30 μM each) for 48 h. One representative blot of 3-4 independent experiments.