Fig. 7

Methazolamide, melatonin, and Trolox activate Nrf2 nuclear translocation in SH-SY5Y cells. SH-SY5Y cells were treated with various concentrations of Aβ42 (0–20 μM) in the presence/absence of MTZ, MEL, or Trolox. Nrf2 nuclear translocation was assessed by incubation with rabbit polyclonal anti-Nrf2 antibody followed by Alexa-488 conjugated anti-rabbit IgG, as detailed in the “Materials and methods” section. Actin was highlighted by immunoreaction with Alexa-588 conjugated phalloidin. Immunofluorescence images depict Nrf2 nuclear signal (green fluorescence) and actin staining (red signal); bar, 25 μm. The graph illustrates the Nrf2 activity in nuclear extracts evaluated by ELISA through the quantitation of Nrf2 binding to immobilized oligonucleotides containing the ARE consensus motif. Bars represent mean ± SEM of triplicate experiments. **p < 0.01 and ***p < 0.001