Fig. 1

TMEM119⁺ CD14⁺ EV release is increased from activated microglia. A qPCR of CD14 expression following LPS treatment of BV-2 microglia. B Western blot of CD14, TMEM119 and TSG101 in EVs isolated from BV-2 microglia supernatant 24 h with or without LPS treatment (500 ng/mL). C Quantification of CD14 expression on EVs isolated from BV-2 microglia supernatant 24 h with or without LPS treatment (N = 3 separate experiments) *indicates p < 0.05 measured using Student’s t-test. D Western blot of TMEM119 and TSG101 on EVs isolated from BV-2 microglia supernatant. E Quantification of TMEM119⁺/CD14⁺ EVs in supernatant 24-h post-LPS treatment measured using nanoflow cytometry. F Immunoprecipitation of TMEM119⁺ EVs out of rat plasma demonstrates co-localization of CD14, CD13, CD9, microglia protein Iba1. G Transmission electron microscopy of unlabeled EVs and immuno-gold mediated labeling of TMEM119 and CD14 on EVs isolated from BV2-microglia 24 h post-LPS treatment. Magnification 46,000 scale bar indicates 200 nm (left), 92,000 (centre and right), scale bar indicates 100 nm