Fig 2: C. albicans induces vesicle release from human blood monocytes.a Vesicle formation (arrow) increases in C. albicans-infected monocytes using confocal laser-scanning microscopy (CLSM). Staining– red: CD63, green: CD14, and orange: nucleic acids after 1 h of co-incubation. Bars: 5 µm. Representative data of five independent experiments are shown. b Release of EVs from the infected monocytes visualized by live cell imaging using CLSM. After phagocytosis (23 min) of C. albicans, the EVs (42 min) (arrows) are released. Staining– green: CD14 and cyan: nucleic acids. Bars: 10 µm. Representative data of three independent experiments are shown. c Infected monocytes in an ex vivo whole-blood model show early (20 min) release of MEVsCa (arrow) by live cell imaging (CLSM). Staining– red: CD63, green: CD14, and orange: nucleic acids. Bars: 10 µm. Representative data of five independent experiments are shown. d Scanning electron microscopy (SEM) confirms the presence of MEVs and MEVsCa released by untreated and infected monocytes (arrows) respectively. Bars: 1 µm. Representative data of three independent experiments are shown.

Fig 3: ABD-engineered EVs bind to HSA in vitro. (A) mCD63-ABD EVs were bound by HSA as detected by widefield imaging of single vesicles. The upper and lower panels indicated the binding of mCD63-Nluc EVs and mCD63-ABD 2nd-Nluc EVs to HSA, respectively. AF647-labelled CD9/CD63/CD81 antibodies cocktail was exploited to detect EVs while AF488-labelled HSA was used to show the co-localization of EVs with HSA. (B) hCD63-ABD EVs were bound by HSA detected by widefield imaging of single vesicles. The binding of hCD63-Nluc EVs and hCD63-ABD 2nd-Nluc EVs with HSA was indicated in the upper and lower panels, respectively. (C) ABD EVs bound to HSA as detected by super-resolution imaging (dSTORM) of single vesicles