Fig. 1

Enrichment and characterization of brain derived extracellular vesicles from MLD mice. A) Schematic of the workflow used to enrich extracellular vesicles (EVs). Total EVs were enriched from brain tissue from MLD and control mice at 30 postnatal days (P30), 3 months (3 m) and 6 months (6 m) of age. B-G) Transmission electron microscopy of isolated EVs. Showing representative imagines obtained from WT (ARSA^+/+) vs MLD (ARSA^−/−) EVs from postnatal day 30 (P30), 3 and 6 months (3 m and 6 m). H) EV samples were subjected to electrophoresis and Western blot analysis for common cellular vesicle markers. Representative images obtained from enriched EVs samples isolated from brains of 6 m revealed little detection of early endosome antigen 1 (EEA1), endoplasmic reticulum calnexin (CALX) and synaptic synaptotagmin 1 protein (SYT1) (n = 1 per genotype and time point). EVs markers flotillin 1 (FLOT1) and ras-related protein Rab-5B (RAB5B) were detected and programmed cell death 6-interacting protein (ALIX) was absent. I) Qualitative analysis by western blot analysis of cell type specific markers showed reactivity for glial fibrillary acidic protein (GFAP, astrocytes), beta-tubulin III (TUBB3, neurons), myelin basic protein (MBP, oligodendrocytes), allograft inflammatory factor 1 (IBA1, microglia) and MHCII. Also, reactivity for Actin and sodium/potassium-transporting ATPase subunit beta-1 (Na + K+ ATPase) were detected (n = 1 per genotype and time point)