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[Preprint]. 2023 Feb 15:rs.3.rs-2551245. [Version 1] doi: 10.21203/rs.3.rs-2551245/v1

Figure 5. Involvement of Serpine-1 in the transfer of Aβ cargo from HBMEC-derived EVs to recipient NPCs.

Figure 5

HBMEC were exposed to HIV (30 ng/ml) and/or 100 nM Aβ (1–40) HiLyte 488 for 48 h, followed by isolation of EVs from the cell culture media and treatment of NPCs for 24 h in the presence or absence of PAI039. (A) Confocal images of recipient NPCs with Aβ HiLyte fluorescence (green) and Mitotracker (red). (B) Dose-dependent PAI039 cytotoxicity in NPCs. (C) Quantification of Aβ HiLyte fluorescence in recipient NPCs. NPCs grown on 96-well plates were exposed to HBMEC-derived fluorescent EVs for 24 h. Controls were exposed to non-fluorescent EVs from HBMEC. Selected NPCs were cotreated with the Serpine-1 inhibitor PAI039 (2 μM) and EVs for 24 h. After washing with PBS, Aβ HiLyte fluorescence was measured (Abs/Em 503/528 nm) in a plate reader. The values were normalized to nuclear DRAQ5 fluorescence. Values are mean ± SEM, n=12–14. One-, two- and three-way ANOVA with Tukey’s multiple comparisons test. *Statistically significant at p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.