Extended Data Fig. 1. In vitro study simulating formation of primary protein corona (1^st corona) and secondary protein corona (2^nd corona) on EVs and liposomes.

(a) EV_2D and (b) Negatively charged liposomes (LIP, as representative of synthetic particles with lipid bilayers) were incubated firstly with albumin labelled with Alexa fluor 488 (ALB-AF488) (1^st corona) then with albumin labelled with Cyanide 5 (ALB-Cy5) (2^nd corona). Fluorescence was evaluated by a plate reader to determine the degree of albumin adsorption. Results confirmed that EV precoating with ALB (1^st corona) induces deposition of more albumin (2^nd corona) on EVs (**p = 0.0015). This is mimicking EVs interaction with blood when injected intravenously. In case of LIP, quenching was observed. Data are presented as mean ± SD (n = 3, biologically independent samples) with two-tailed unpaired t-Test analysis. The figure was created with BioRender.com.

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