Fig. 4

(a) Detection of drug-resistant proteins on the surface of HCC1395, MDA-MB-231, and MDA-MB-468 cells under different conditions. MFI for each target is shown on the y-axis: untreated control cells (black), wild-type cells (red), stable drug-resistant cells (green), drug-resistant cells after 48 h of AC treatment (orange), and drug-resistant cells after 48 h of T treatment (blue). Data are shown as mean ± SD of three independent experiments. Significant differences between groups were determined using one-way ANOVA (*p < 0.05; **p < 0.01; and ***p < 0.001). (b) TEM images of EVs extracted from each cell. Black scale bars represent 100 nm. (c) A representative confocal image of enriched EVs by immuno-beads with a red box highlighting a SEM image of EVs attached to immuno-beads. White scale bars represent 10 μm, except for the SEM image which represents 200 nm. (d) Flow cytometry gating strategies for drug-resistant EV markers (d) Comparison between wild-type and drug-resistant clones in cell lines and EVs. Significant differences between groups were determined using one-way ANOVA (ns, not significant; *p < 0.05; **p < 0.01; and ***p < 0.001)