Fig. 8

RBC-EVs impair astrocytic protection of neurons. a Schematic representation demonstrating the setup of the trans-well model. Astrocytes were cultured in the insert compartment of the trans-well and neurons were cultured in the bottom of the wells. b Representative images of cultured neurons labelled with synaptophysin. Note that RBC-EV pre-treated astrocytes cannot prevent the loss of synaptophysin-positive signals in the presence of glutamate treatment (Scale bar, 4 μm). c-d Quantification analysis of number synaptophysin puncta (means + S.E.M; n = 3; *p < 0.05 by One-way ANOVA). e Quantification analysis of fluorescence intensity of synaptophysin labeling in cortex (CTX), striatum (STR), midbrain (MIDB) and cerebellum (CERE) (means + S.E.M; n = 5; ****p < 0.0001 by One-way ANOVA test). f-g Graphs show the levels of oligomeric α-syn in synaptosomes of mouse cortex (CTX), striatum (STR), midbrain (MIDB) and cerebellum (CERE) after chronic RBC-EV injection (f) and normalized value against the concentration of oligomeric α-syn in cerebellum (g) (n = 5 independent animals were used in each group; means + S.E.M; *p < 0.05 by Student’s t-test *p < 0.05 compared to control-EVs injected group). h Quantification analysis of fluorescence intensity of synaptophysin labeling in cortex (CTX), striatum (STR), substantia nigra (SN) and cerebellum (CERE) of healthy control or PD human postmortem tissues (means + S.E.M; n = 5; *p < 0.05 by One-way ANOVA test)