Fig. 4

Uptake of intracranially injected glioma-derived fluorescent EVs is associated with a decrease in microglial sensing capability. a Schematic overview of EV isolation from glioma cells in culture using differential centrifugation. b Size distribution analysis using NTA of isolated EVs shows small and larger vesicles present in the EV preparation. c Western blot analysis shows GFP present in cells and EV, and extracellular vesicles markers (ALIX, TSG101 and Flotillin-1) enriched in vesicles lysate and GAPDH is detected in cellular lysate only. d Microglia were identified as CD11b^high/CD45^med cells (blue gate). Microglia were then sorted based on the GFP signal detected as the upper limit in control. In mice injected with GL261.BpalmGFP EVs, a population of GFP-positive microglia was identified (green gate in the GFP/RFP plot). e Heatmap of sensome genes ordered top to bottom by highest up- to downregulated for mouse EV-GFP^pos tumor microglia compared to wildtype (same order as Fig. 3b). Similar patterns are observed for genes up- and downregulated compared to the mouse tumor-derived profile. Asterisk (*) indicates significant (multiple testing adjusted p value < 0.05) differential expression. Error bar represents the SEM, bar represents the mean, and dots display individual measurements (n = 3)