Figure 3. In SH-SY5Y cells Nrf2 shRNA or KO abolishes CBR-470-1-induced cytoprotection against MPP+. Expression of Nrf2 mRNA in stable SH-SY5Y neuronal cells with Nrf2 shRNA (“sh-Nrf2”) or a lenti-CRISPR/Cas9-Nrf2 KO construct (“ko-Nrf2”), as well as in the parental control cells (“Pare”), was shown (A); Cells were treated with CBR-470-1 (“CBR”, 10 μM) or the vehicle control (“Veh”) for applied time periods, expression of listed mRNAs and proteins was shown (B–D); Alternatively, cells were pre-treated for 2h with CBR-470-1 (“CBR”, 10 μM) or the vehicle control (“Veh”), followed by MPP+ (3 mM) stimulation for 48h, cell viability and death were tested by CCK-8 (E) and medium LDH release (F) assays, respectively. Expression of listed proteins was quantified and normalized to the loading control (B, D). Bars stand for mean ± standard deviation (SD, n=5). * P< 0.05 (E, F). #P< 0.05 vs. “Pare” cells of same treatment (A, C, E, F). Experiments in this figure were repeated four times, with the similar results obtained.