Journal: PNAS Nexus
Article Title: Monitoring drug metabolic pathways through extracellular vesicles in mouse plasma
doi: 10.1093/pnasnexus/pgae023
Figure Lengend Snippet: Comparison between ultracentrifugation (100,000 × g ) and EVtrap for liver tissue EV isolation. Particles were isolated from 0.2 mL supernatant after 10,000 × g centrifugation on tissue homogenate. A) Western blotting data showing programed cell death six-interacting protein (Pdcd6ip, also known as Alix) and heat shock protein family A member 8 (Hspa8) as EV markers, as well as Calnexin (Canx) as contaminant protein (negative control). B) Nanoparticle analysis was performed by TRPS on 100,000 × g pellets or eluates from EVtrap beads. Relative abundances of representative C) EV markers and D) contaminant proteins by LC–MS/MS analysis with the z -score color indicated, respectively. 0.5 μg resulting peptides from each method were loaded and technical triplicates were employed. E) The numbers of identified EV proteins by ultracentrifugation and EVtrap isolation methods. Selected proteins were identified in at least two out of three technical replicates.
Article Snippet: EVtrap kits, including EVtrap beads as a suspension in water, were provided by Tymora Analytical Operations and were used as described before ( , , ).
Techniques: Comparison, Isolation, Centrifugation, Western Blot, Negative Control, Liquid Chromatography with Mass Spectroscopy