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Figure 3. Preparation and validation of transferrin-conjugated EVs. (A) Schematic illustration of the preparation of functionalized tt-Mfn-EVs. This illustration was created using BioRender.com; (B) Dot blot images of activated transferrin conjugated DSPE lipid (Tf-DSPE), empty EVs, and Tf-DSPE-treated EVs loaded with GNP-CDDP (tt-Mfn-EVs); (C) MS/MS graph of transferrin peptide sequence in Tf-DSPE-treated EVs; (D) Quantification of transferrin peptide sequence on EVs and EV-Tf. The bar graph represents the mean ± SD from three replicates (n = 3). Statistical significance was assessed using an unpaired Student’s t-test, with P values indicated as ^*P < 0.05. Created in BioRender. Ramesh, R. (2025) https://BioRender.com/yup6by2. EVs: Extracellular vesicles; tt-Mfn-EVs: tumor-targeted multifunctional extracellular vesicles; DSPE: 1,2-distearoyl-sn-glycero-3-phosphoethanolamine; GNP: gold nanoparticle; CDDP: cisplatin; MS: mass spectrometry; SD: standard deviation.