4.9. Determination of Nrf2 Nuclear Translocation

For the separation of nucleic extracts from whole lysates, the NE-PER kit was used according to the manufacturer’s instructions. Briefly, incubated SH-SY5y cells under the indicated conditions were harvested and lysed using lysis buffer I. After centrifugation with lysis buffer II, the lysate was removed for further analysis (cytosolic extract). Pellets were lysed using lysis buffer III and centrifuged to isolate the nucleic extracts. Approximately 30 μg of lysate was loaded onto 6% SDS-PAGE gels. The expression of LaminB and β-actin indicates the purity of the nucleic and cytosolic extracts, respectively. The detected Nrf2 was normalized to the expression of LaminB (nucleic extract) and β-actin (cytosolic extract).