Fig. 8

Alix and Syntenin-1 knockdown ameliorates the neurotoxicity conferred by APP^swe-containing EVs. a Differentiated primary neurons were stained with anti-Map2 to demonstrate neuronal origin. Scale bar = 50 μm. b Neurons were treated with puromycin, purified oligomeric amyloid beta peptide, or EVs derived from control, Alix shRNA, or Syntenin-1 shRNA cell lines expressing APP^Swe. Neuronal cells were stained with both Annexin V (green) and propidium iodide (PI; red) to assess for levels of apoptosis and cell necrosis, respectively. Scale bar = 100 μm. c Cell death was quantitated by Annexin V and PI puncta per microscopic field. A minimum of 200 cells were counted per group. Imaging was performed using a Keyence BZ-X710 fluorescence microscope. Images are representative across three independent experiments. One-way ANOVA with post hoc Tukey’s multiple comparison test: ***, p < 0.001; **, p < 0.01; *, p < 0.05