Fig. 3
From: Oxidized SOD1 accelerates cellular senescence in neural stem cells
EVs from senescent NSCsWT contain misfolded and oxidized hSOD1. A. Level of misfolded hSOD1 in EVs isolated from P3 and P5 NSCsWT and NSCsG93A as detected by affinity binding assay with the GST-tagged CT4. Blots were probed with an anti-hSOD1 antibody. Input: total SOD1 in each lysate. Full-length blots/gels are presented in Additional file 1: Fig. 3A. B. EM analysis revealed the presence of EVs by negative staining. Scale bar: 100 nm. C. Thiol-disulfide status of the SOD1 in EVsWT and EVsG93A from indicated passages of NSCs. The thiol content was normalized to total hSOD1. Two-way ANOVA followed by Sidak’s multiple comparisons test was performed. D. SOD1 expression in EVs from P5 NSCsWT and NSCsG93A was analyzed by non-reducing SDS-PAGE with antibodies to hSOD1 and misfolded hSOD1 (A5C3). Full-length blots are presented in Additional file 1: Fig. 3D. The experiments were done in triplicate from three independent cultures.