Figure 6.

A differential centrifugation approach allows the isolation of EVs subpopulations from NRK52E conditioned medium. (A) Flowchart of the differential centrifugation steps for the isolation of EVs. P1–P4 indicate pellets, and S1–S4 indicate supernatants. (B) Analysis of microparticle size distribution in fractions P3 and P4 was performed by nanoparticle tracking analysis with qNano (Izon). Ectosomes were recovered from the P3 fraction, and exosomes were recovered from the P4 fraction. Representative particle size distributions of an ectosome-enriched 10,000-g pellet (dark gray) and an exosome-enriched 110,000-g pellet (light gray). (C) Expression of FLOT2 was measured by Western blotting in equal amounts from the differential centrifugation fractions obtained from the conditioned medium of NRK52E cells cultured in serum-free conditions.