Fig. 6
From: Oxidized SOD1 accelerates cellular senescence in neural stem cells
Blocking misfolded SOD1 reduces SOD1 aggregation and mitigates the neurotoxicity of EVs isolated from senescent NSCs. A. and B. Blocking misfolded SOD1 reduces SOD1 aggregation. P5 EVsWT were incubated for 2 h with or without the CT4 motif before adding it to P3 NSCsWT. After 24 h, protein samples were separated into detergent-insoluble and soluble fractions and analyzed by Western blot with an anti-hSOD1 antibody. Full-length blots/gels are presented in Additional file 1: Fig. 6A. Total protein generated by stain-free visualization was used to ensure equal loading. Two-way ANOVA followed by Sidak’s multiple comparisons test was performed. C. Neurotoxicity of P5 EVs to P3 NSCsWT. Cells were immunostained with antibodies to misfolded hSOD1 (B8H10, Red) and MAP2 (Green). Scale bar: 50 μm. D. Quantification of MAP2 positive neurons. E. Release of lactate dehydrogenase. For D and E, one-way ANOVA followed by Bonferroni’s Multiple Comparison Post-Test was performed. The experiments were done in triplicate from three independent cultures