Fig. 4

BG-EVs are taken up by astrocytes: we incubated 10,000 astrocytes from CX3CR1^+/GFP mice with increasing concentrations (0, 12.5, 25, 50, 100 and 200 µg/mL) of SytoSELECT labeled BG-EVs in the presence of NucBlue (a live cell stain, 30 µL/mL) and seeded in a glass-bottom 96 well plate 24-h prior to experiment. Kinetics imaging (every 3 h) was recorded over the course of 18 h using automated Lionheart FX software [18] (Biotek) after which total GFP (ex/em) and Nucblue (ex/em) intensity were recorded using a plate reader (Synergy H1 Biotek). Astrocytes were assessed for viability using Cell Titer Glow assay (Promega). A Representative 10 × images at 18 h timepoint after addition of BG-EVs. B Total gfp and nucblue intensity as measured by the plate reader. E Viability of astrocytes treated with BG-EVs. Ordinary one-way ANOVA multiple comparison test (Tukey’s test) was used to assess statistical differences. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, non-significant. Error bars represent Standard Deviation