Figure 5

Characterization of HLSC-derived EVs surface markers and NTA analyses. (A) Nanosizer analysis of purified HLSC-derived EVs: curve 1 shows the relationship between particle distribution (left Y axis) and particle size (X axis); curve 2 shows the correlation between cumulative percentage distribution of particles (percentile in right Y axis) and particle size (X axis). Mean size and particle concentration values were calculated by Nanoparticle Tracking Analysis (NTA). (B) Representative Western blot analysis of exosomal markers CD63, CD81, Alix and Hsp90 expressed by HLSCs and derived EV (10 μg protein/well). (C-D) Representative FACS analyses of the expression of classical mesenchymal (C) and exosomal markers (D) by HLSC-derived EVs. Dot lines indicate the isotopic controls. Three different EV preparations were tested with similar results. EVs, extracellular vesicles; FACS, fluorescence-activated cell sorting; HLSC, human liver stem cell.