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Impact of EVs derived from CSC-like MDA-MB-231 cells on MCF-7 cells. (A) Uptake of PKH67-labeled EVs in MCF-7 cells. Left to right: brightfield, DAPI nuclear stain, PKH67-labeled EVs, and merged. Scale bar: 25 µm. (B) miRNA expressions of stemness-related markers in MCF-7 cells after 24 h incubation with PBS control (PBS), static cell-derived EVs (static), and FSS-derived EVs (FSS). Expressions were normalized to reference miRNA marker miR-30e. n = 3 biological replicates each with 2 or 3 technical replicates; mean ± standard error; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns = not significant via one-way ANOVA. (C) Cancer stem cell-like subpopulation (CD44⁺/CD24⁻; lower right quadrant) of MCF-7 cells after 24 h of EV introduction measured via flow cytometry (n = 3).

Impact of EVs derived from CSC-like MDA-MB-231 cells on MCF-7 cells. (A) Uptake of PKH67-labeled EVs in MCF-7 cells. Left to right: brightfield, DAPI nuclear stain, PKH67-labeled EVs, and merged. Scale bar: 25 µm. (B) miRNA expressions of stemness-related markers in MCF-7 cells after 24 h incubation with PBS control (PBS), static cell-derived EVs (static), and FSS-derived EVs (FSS). Expressions were normalized to reference miRNA marker miR-30e. n = 3 biological replicates each with 2 or 3 technical replicates; mean ± standard error; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns = not significant via one-way ANOVA. (C) Cancer stem cell-like subpopulation (CD44⁺/CD24⁻; lower right quadrant) of MCF-7 cells after 24 h of EV introduction measured via flow cytometry (n = 3).

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