EV Characterization
(A) NanoSight representative images of uEVs isolated by ultracentrifuge (uEV) and floating (uEV float) protocols and of MSC EVs. (B) Representative western blot showing renal and exosomal markers AQP1, AQP2, Klotho, CD63, and calreticulin in uEVs, uEVs float, and MSC EVs; calreticulin expression by fibroblast cells was analyzed as positive control. Three different uEV preparations were analyzed with similar results. (C) Representative micrographs of transmission electron microscopy obtained from purified uEVs (scale bars: right panel, 50 nm; left panel, 200 nm). (D) Representative cytofluorimetric analyses of uEVs showing the positive expression of renal and exosomal markers AQP1, AQP2, PDX, CD24, CD81, CD63, and the negative expression of monocyte, platelet, and endothelial markers CD45, CD42b, and VEGFR2 (VR2), respectively. Dotted line histograms specify isotypic controls. Three different uEV preparations were analyzed with similar results.