Fig. 5.
Stable overexpression of Nrf2 enhances resistance of SH-SY5Y neuroblastoma cells to chemotherapeutic agents. (A) Two SH-SY5Y-derived cell lines stably expressing the control vector or HA–Nrf2 were established using a lentivirus-based approach. Relative amounts of mRNAs for Nrf2, NQO1 and HO-1 were measured (top three panels of first row). The elevated protein levels of Nrf2 and NQO1 were confirmed by immunoblot analysis (left panels of second row). The NQO1 enzymatic activity and the intracellular glutathione level were measured (middle and right panels of second row). (B) Cell viability of these two cell lines in response to 48 h cisplatin treatment was measured by the MTT assay. (C) Apoptotic cell death in response to cisplatin treatment was also measured using Cell Death Detection ELISAPLUS. (D) Following cisplatin treatment, cells were labeled with fluorescein-deoxyuridine triphosphate using the In Situ Cell Death Detection kit (left panel). The number of apoptotic cells was counted under fluorescent microscope and the average number of apoptotic cells per field is listed in the table. The data are presented as means ± SDs. *P < 0.05.