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. 2023 Dec 10;12(12):12392. doi: 10.1002/jev2.12392

FIGURE 4.

FIGURE 4

Bodipy sEV separate as a single peak enriched in tetraspanins. (a) Bodipy sEV 100K pellets were loaded at the bottom of an iodixanol density gradient and ultracentrifuged for 16 h. Fractions were collected from the top of the gradient and were analyzed for Bodipy sEV number by FC before and after the addition of Octyl‐beta‐glucoside (OG). Data are expressed as mean ± SEM (n = 14). (b) 100K pellets from conditioned media of untreated Mel501 cells were incubated with CFSE to non specifically label the total population of sEV and loaded at the bottom of an iodixanol density gradient. Data are expressed as mean ± SEM (n = 4). (c) Density of the fractions was measured by refractometry. Data are expressed as mean ± SEM (n = 17). (d) The total volume of each fraction was analyzed by Western blot. Data shown are representative of three independent experiments. (d) Analysis of colocalization of tetraspanins fluorescent antibodies with Bodipy sEV.