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Figure 3. Overview of EV Isolation. (A) Isolation of MEVs. The process involves ultracentrifugation of pretreated samples, preceded by low-speed centrifugation to remove impurities such as cells and cellular debris; (B) Isolation of PEVs. After pretreatment, plant juice undergoes a sequential centrifugation process at low, medium, and high speeds. PEVs are then obtained from the liquid layer using the discontinuous sucrose gradient method, as shown in the figure; (C) Isolation of BEVs. Samples are first centrifuged at low speed and filtered to remove bacteria. The filtered supernatant is then concentrated using ultrafiltration membranes. BEVs are isolated by two rounds of ultracentrifugation and finally resuspended in PBS. Created in BioRender. Bigbone B. (2025) https://BioRender.com/zavrv7g. EVs: Extracellular vesicles; MEVs: mammalian extracellular vesicles; BEVs: bacterial extracellular vesicles; PEVs: plant-derived extracellular vesicles; UC: ultracentrifugation; DGC: density gradient centrifugation; UF: ultrafiltration; PBS: phosphate-buffered saline.