Fig. 4
Effect of Intercept-platelet lysates on LUHMES cell viability. LUHMES cells were seeded at a density of 2.5 × 105 cells/well. Following day five of differentiation, the cells were treated with 5% Intercept-platelet lysate (v/v) for 24 or 48 h. Controls were cells grown in differentiation medium only. Cells treated with Erastin only were used as a positive control for lipid peroxidation evaluation. After 24 h of incubation, the cells were collected and stained with propidium iodide for cell viability analysis or C11-Bodipy for lipid peroxidation measurement using flow cytometry. After 48 h of incubation, cell viability was assayed using cell counting kit-8 (CCK-8 test). Timeline of experiments is given in (a). b and c cell viability after 24 and 48 h of incubation, respectively. d lipid peroxidation levels. For proteins expression evaluation, the lysates of treated-cells were subjected to Western blot analysis. e Representative Western blots for Tyrosine hydroxylase (TH) and Neuron Specific Enolase (NSE) expression. f and g Quantifications. All data (at least n = 3) were expressed as the mean ± SD of untreated controls. ns: not statically significant; *p < 0.05, **p < 0.01 ***p < 0.001 vs. untreated controls using One-way analysis of variance (ANOVA) followed by Fisher’s Least Significant Difference (LSD) test. Abbreviations: day 0 (d0); heat-treated platelet pellet lysate (HPPL Intercept-platelet pellet lysate (I-PPL); heat-treated Intercept-platelet pellet lysate (I-HPPL)