Figure 5. SphK1 knockout intensifies MPP+-induced neuronal cell death. Stable SH-SY5Y cells expressing the lentiCRISPR-GFP-SphK1-KO construct (“koSphK1” cells) were further transduced with or without the lentiviral construct encoding the anti-sense of premiR-6862 (lv-antagomiR-686), control cells were transduced with lentiCRISPR-GFP empty vector (“Cas9-C”); Cells were treated with or without MPP+ (3 mM) and then cultured for applied time periods, SphK1 mRNA and protein expression was tested (A, B); Cell viability and death were tested by CCK-8 (C) and LDH release (D) assays, respectively; Cell apoptosis was examined by nuclear TUNEL staining assay (E). miR-6862 expression was shown (F). Bars stand for mean ± standard deviation (SD, n=5). * P < 0.05 vs. “Ctrl” treatment in “Cas9-C” cells. #P < 0.05 vs. MPP+ treatment in “Cas9-C” cells. “n.s.” stands for non-statistical difference. Experiments in this figure were repeated five times, with the similar results obtained.