Fig. 6
Neuroprotective ability of Intercept-platelet lysates. LUHMES cells were seeded at a density of 2.5 × 105 cells/well. After day five of differentiation, cells were pre-treated with 5% Intercept-platelet lysate (v/v) for 1 h prior to the addition of Erastin and incubated for either 24 or 48 h. Controls (untreated) cells were grown in differentiation medium and treated, or not, with Erastin. 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. 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. The protective effect of Intercept-platelet lysate was also evaluated using primary neurons culture. At DIV21, the mature neuronal culture was stimulated with 0.5% Intercept-platelet lysate 1 h prior to the addition of 1.25 μM Erastin. General cytotoxicity was evaluated by LDH assay (h). *p < 0.05, **p < 0.01 ***p < 0.001 vs. untreated controls, ####p < 0.0001 vs erastin treated cells using One-way analysis of variance (ANOVA)