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Figure 4 | Molecular Brain

Figure 4

From: Exosomes neutralize synaptic-plasticity-disrupting activity of Aβ assemblies in vivo

Figure 4

Exosomal surface proteins are essential for the protective role of exosomes against the synaptic-plasticity-disrupting action of ADDLs. (A) Trypsinization (T+) was effective in removing surface proteins (PrPC and CD81), but not luminal protein (Alix) of N2a cell-derived exosomes when compared to non-trypsinized (T-) control. Top, representative blots and bottom, normalized mean O.D. of immunoblots, n = 5. (B) Trypsinized exosomes (T+ Exo, red line) had similar sizes (74 ± 3 nm), compared with those of non-trypsinized exosomes (T- Exo, black line) (83 ± 7 nm, P > 0.05, n = 3), as measured by DLS. (C) By EM, the structure and size of T+ Exo were indistinguishable from T- Exo (Scale bar: 100 nm). (D) T+ Exo (5 μl, asterisk) was unable to prevent the plasticity-disrupting-activity of ADDLs (10 pmol in 5 μl, hash), but which can be neutralized by T- Exo. An arrow indicates HFS application and insets show representative traces at the color-matched time points. Calibration: 1.5 mV and 10 ms. (E) Lack of effect of i.c.v. injection (asterisk) of T+ Exo (n = 4) on the stability of fEPSP recordings. (F) Comparing to T- Exo, T+ Exo showed a reduced ability to sequester ADDLs. Left panel: representative immunoblot using antibodies against Aβ (6E10), Alix and Flotillin-1 (P, pellet; S, supernatant). Right panel: mean % of ~12 and 16 kDa Aβ bound to exosomes (P) relative to total ~12 and 16 kDa Aβ (P + S). Error bars, ± SEM. Statistical significance was expressed as *, P < 0.05; **, P < 0.01.

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