Figure 4.

Impact of PC10:0/10:0, PC20:0/20:0 or solvent on the activity of antioxidative enzymes and on catalase gene expression. Cells were incubated with the solvent EtOH (0.2%) (set as 100%), PC10:0/10:0, PC20:0/20:0 (10 µM) for 18 h in absence (a,b,e) or presence of PPARγ antagonists (c) and inhibitors of protein biosynthesis (d). Catalase, GPx and SOD activity in the homogenates of (a) SH-SY5Y cells (catalase: n ≥ 5, GPx: n ≥ 5, SOD: n ≥ 6) and (b) Neuro2a cells (catalase: n ≥ 6, GPx: n ≥ 5, SOD: n ≥ 6) determined by using the corresponding enzyme activity assay kits. The activity level of the respective enzyme in untreated cells is indicated by a dotted line. H₂O₂ released by SH-SY5Y cells measured by using Amplex Red (5 µM) and HRP (0.01 U/ml) after treatment with phospholipids or solvent along with (c) 5 µM BADGE / 5 µM GW9662/ DMSO (n ≥ 4) and (d) 20 µM cycloheximide/ 2 µM puromycin/ DMSO (n ≥ 5). (e) Catalase gene (CAT) expression in SH-SY5Y cells measured by RT-PCR using two different CAT primer pairs and two different house keeping genes (ACTB: β-actin, TBP: TATA-binding protein) for normalization (n = 7). Error bars represent SD (a,b,e) or SEM (c,d). Asterisks show the statistical significance calculated by one-way ANOVA followed by post hoc testing using Tukey's test (** p ≤ 0.01 and *** p ≤ 0.001). Figure was created using Origin Pro 2020b and CorelDRAW Graphics Suite 2020.