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Figure 4. Characterization of the EVs from UF-SEC-UC and UC-SEC-UC in pbMEC and MAC-T (A, B) electron microscopy picture of a vesicle from UF-SEC-UC (A) and UC-SEC-UC (B) on pbMEC conditioned medium. (C, D) Total number of particles per 10 mL of conditioned medium (C) and size ranges (D) from EVs from UF-SEC-UC and UC-SEC-UC in pbMEC conditioned medium. The TRPS was performed with a NP100 nanopore. (E) Protein concentration of the single fractions from SEC from pbMECs conditioning medium. (F, G) Electron microscopy pictures of vesicles from UF-SEC-UC (F) and UC-SEC-UC (G) on MAC-T conditioned medium. (H, I) Total number of particles per 10 mL of conditioned medium (H) and size ranges (I) from EVs from UF-SEC-UC and UC-SEC-UC in pbMEC conditioned medium. The TRPS was performed with a NP100 nanopore. (L) Western blot of pbMECs and MAC-T EVs pellet. Whole cells lysates were used as a positive control for calnexin, while EVs from milk were used as a positive control for EV markers. Full-length blots are presented in Supplementary Figure 2. In (C-E, H, I), values are mean ± SD of three replicates. Wilcoxon test (C, H) was performed. The differences were considered significant when P < 0.05. FBS: Fetal bovine serum; EVs: extracellular vesicles; UC: ultracentrifugation; SEC: size exclusion chromatography; pbMEC: primary bovine mammary epithelial cells; UF: ultrafiltration.