Fig. 1: Detection and characterization of LRRK2 and Rab epitopes in urinary exosome enriched extracellular vesicles (uEVs). | npj Parkinson's Disease

Fig. 1: Detection and characterization of LRRK2 and Rab epitopes in urinary exosome enriched extracellular vesicles (uEVs).

From: Alterations in the LRRK2-Rab pathway in urinary extracellular vesicles as Parkinson’s disease and pharmacodynamic biomarkers

Fig. 1

a Flow diagram of the differential centrifugation protocol employed to isolate uEVs. The full method is described in detail in materials and methods. b Nanoparticle tracking analysis of extracellular vesicles isolated from urine according to the method schematized in (a) (sample P3). c LRRK2 detection via immunogold labeling in negative stain transmission electron microscopy (TEM). The panel shows a TEM image of uEVs and dual immunogold labeling with anti-LRRK2 antibodies (RabMab UDD3 and Neuromab N241, revealed by large and small gold particles, respectively) illustrating LRRK2 labeling in uEVs. Scale bar, 100 µm. d Western blot detection of total and phospho-epitopes of LRRK2 with multiple different antibodies, using different fractions of urine (pellet fractions P1, P2, and P3 as schematized in A) as well as recombinant LRRK2. The presence of the exosome marker TSG101 in P3 indicates that this fraction is enriched in exosomes.

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