fig1

Figure 1. Intravesicular protein detection by dSTORM. (A) Confocal micrographs of FEMX-I and GFP⁺ FEMX-I cells stained with phalloidin (actin, red) and DAPI (blue). (B) Size and concentration of EVs derived from cells in panel A were determined by NTA. (C) Cells and EVs were solubilized and subjected to Western blot analysis. Note the absence of GFP in the parental cells and EVs and of calnexin (CNX) in both EV samples. (D) EVs derived from GFP⁺ FEMX-I were labeled with DiI dye and analyzed by confocal microscopy. (E and F) EVs were permeabilized and labeled for CD9 or CNX (blue), CD63 (yellow), and GFP or CD81 (red) and analyzed by dSTORM then quantified. (G) EVs derived from GFP⁺ FEMX-I were subjected to increasing concentrations of Triton X-100, analyzed by dSTORM, and quantified. For all EV quantification, a range of 2,000 to 3,000 EV particles were analyzed per replicate per condition. N.s.: not significant; ***: P < 0.001; Scale bar: 10 µm (A, D) or 100 nm (E).