Research Paper Volume 12, Issue 13 pp 13388—13399

Figure 4. PGK1 shRNA or KO activates Nrf2 signaling and inhibits MPP⁺-induced cytotoxicity in SH-SY5Y cells. Expression of listed mRNAs and proteins in the stable SH-SY5Y neuronal cells with PGK1 shRNA (“sh-PGK1”), the lenti-CRISPR/Cas9-PGK1 KO construct (“ko-PGK1”), as well as in the parental control cells (“Pare”), was tested by qPCR (A, D) and Western blotting (B, E) analyses, with the relative ARE luciferase activity tested as well (C); Alternatively, cells were treated with MPP⁺ (3 mM) for 24-48h, single strand DNA contents (F) and mitochondrial depolarization (JC-1 green fluorescence intensity, G) were tested; Cell viability, death and apoptosis were tested by CCK-8 (H), medium LDH release (I) and nuclear TUNEL staining (J) assays, respectively. Expression of listed proteins was quantified and normalized to the loading control (B, E). Bars stand for mean ± standard deviation (SD, n=5). * P< 0.05 vs. “Pare” cells (A, C, D). * P< 0.05 vs. “Pare” cells with “Ctrl” treatment (F–J).^#P< 0.05 vs. MPP⁺-treated “Pare” cells (F–J). Experiments in this figure were repeated four times, with the similar results obtained.