Cholesterol impairs the binding of sEVs to laminin and the MRC-5 cell PM. (A) Fluorescence images of sEV–CD63Halo7-SF650T particles bound to laminin (LN) on glass before and after cholesterol depletion by MβCD and the numbers of attached sEVs per image (82 × 82 μm). The cholesterol content in PC3 cell–derived sEVs was reduced to 16% after treatment with MβCD. (B and C) The numbers of sEV–CD63-Halo7-SF650T particles attached to glass coated with laminin before and after treatment with saponin (B) and the addition of cholesterol by MβCD–cholesterol complex (C).The cholesterol content was increased to 186% after treatment with the MβCD–cholesterol complex. (D) Fluorescence images of an MRC-5–GFP cell and sEV–CD63Halo7-SF650T particles on the MRC-5 cell after 30 min of incubation. (E and F) Time course of the number of sEV–CD63-Halo7-SF650T particles per 1,000 μm2 attached to the MRC-5 cell membrane before and after treatment with MβCD (n = 16 cells) (E) or the MβCD–cholesterol complex (n = 8 cells) (F). Data are presented as the mean ± SE. n.s., nonsignificant difference; **P < 0.01; ***P < 0.001 according to Welch’s t test (two-sided).
